CN114015775A - Molecular probe for treating head and neck squamous cell carcinoma and application thereof - Google Patents
Molecular probe for treating head and neck squamous cell carcinoma and application thereof Download PDFInfo
- Publication number
- CN114015775A CN114015775A CN202111325700.4A CN202111325700A CN114015775A CN 114015775 A CN114015775 A CN 114015775A CN 202111325700 A CN202111325700 A CN 202111325700A CN 114015775 A CN114015775 A CN 114015775A
- Authority
- CN
- China
- Prior art keywords
- tcab1
- selenium compound
- neck squamous
- head
- atr
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57407—Specifically defined cancers
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/10011—Adenoviridae
- C12N2710/10041—Use of virus, viral particle or viral elements as a vector
- C12N2710/10043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/21—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Pseudomonadaceae (F)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- General Engineering & Computer Science (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a molecular probe for treating head and neck squamous cell carcinoma and application thereof, and relates to the technical field of head and neck squamous cell carcinoma treatment. According to the molecular probe for treating head and neck squamous cell carcinoma, the influence of a selenium compound on the stability of a head and neck squamous cell carcinoma cell genome is researched: firstly, researching the function of activating the ATR channel by a selenium compound; secondly, analyzing the action of the selenium compound on TCAB1 transcriptional inactivation; the interaction of the ATR pathway and the TCAB1 pathway was investigated; finally, analyzing the feasibility of the genome stability intervention treatment of the head and neck squamous cell carcinoma under the action of the selenium compound; during research, the tumor cell genome comprises two human oral squamous carcinoma cell lines (HSC-3, Cal-27), a nasopharyngeal carcinoma cell line CNE1 and an adenoid cystic carcinoma cell line ACC2, and the acting reagents of the selenium compound are sodium selenite and seleno-L-methionine. Aiming at the research focus of selenium anticancer, the invention aims at achieving accurate targeted therapy by analyzing the node and mechanism of mutual regulation and control of an ATR passage and telomerase TCAB1 under the action of a selenium compound, and can possibly obtain a more obvious anticancer effect.
Description
Technical Field
The invention relates to the technical field of head and neck squamous cell carcinoma treatment, in particular to a molecular probe for head and neck squamous cell carcinoma treatment and application thereof.
Background
The incidence of head and neck malignant tumors is 6 th of the whole body malignant tumors, wherein more than 90 percent of the head and neck malignant tumors are squamous cell carcinoma, and although the treatment method is continuously improved in the last 20 years, the five-year survival rate is still about 50 percent, and no effective measures are provided for the high recurrence rate, the high metastasis rate and the chemoradiotherapy resistance of head and neck squamous cell carcinoma clinically at present.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a molecular probe for treating head and neck squamous cell carcinoma and application thereof, and solves the problem that no effective measure exists for high recurrence rate, high metastasis rate and chemoradiotherapy resistance of head and neck squamous cell carcinoma clinically.
(II) technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme: the molecular probe for treating the head and neck squamous cell carcinoma and the application thereof are characterized in that the influence of a selenium compound on the genome stability of head and neck squamous cell carcinoma cells is researched:
researching the action of activating an ATR (attenuated total reflectance) channel by a selenium compound;
analyzing the action of the selenium compound on TCAB1 transcriptional inactivation;
step three, researching the interaction of the ATR channel and the TCAB1 channel;
analyzing the feasibility of the genome stability intervention treatment of the head and neck squamous cell carcinoma under the action of the selenium compound;
during research, the tumor cell genome comprises two human oral squamous carcinoma cell lines (HSC-3, Cal-27), a nasopharyngeal carcinoma cell line CNE1 and an adenoid cystic carcinoma cell line ACC2, and the acting reagents of the selenium compound are sodium selenite and seleno-L-methionine.
Preferably, the specific research method in the step one is as follows:
a. systemic analysis of molecular expression of ATR pathway signaling in selenium compound-treated head and neck squamous carcinoma cells: detecting the capability of forming foci of ATRIP, TopBP1, 9-1-1 and Mre11-Rad50-Nbs1 complex, H2AX, CTIP and PCNA signal molecules by immunofluorescence and immunoblotting;
b. formation of single-stranded DNA intermediate: detecting the forming capacity of single-stranded DNA (ssDNA) in the selenium compound processing head and neck squamous carcinoma cells by using a phosphorylation RPA and non-denaturation BrdU immunofluorescence method;
stability of dna replication forks: BrdU and IdU are used for labeling cells in an interactive mode, and the length of an IdU single-color fluorescence labeling area is observed after ultraviolet rays or camptothecin are used for treating head and neck squamous carcinoma cells so as to indicate the capacity of restarting and extending a replication fork;
d. screening of selenium compound acting protein: constructing an adenovirus vector, expressing selenium compound related protein with a TAP label, preparing a high-titer adenovirus vector, infecting head and neck squamous carcinoma cells (108 cell numbers), extracting total protein or nucleoprotein, specifically adsorbing a selenium compound TAP and an interaction protein complex by using magnetic beads (Dynal beads) which are covalently bonded with rabbit IgG, separating and cutting a specific protein band by SDS-PAGE electrophoresis, then carrying out MALDI-TOF or liquid chromatography-related mass spectrometry, identifying polypeptide fingerprints, and screening target protein related to an ATR (attenuated total reflectance) channel.
Preferably, the specific research method in the second step is as follows:
a. influence of selenium compound on contents of components of telomerase complex and localization of Cajal body: TRAP-ELISA for detecting telomerase activity; detecting telomere length by southern blot; westernblot detection of TCAB1, hTERT and dyskherin; notherblot detection of hTR levels; using antibodies of TCAB1, p80-coilin (a marker protein of CB), dyskerin and hTERT to carry out a co-immunoprecipitation experiment to detect the interaction of TCAB1 with dyskerin, hTERT and CB under the action of a selenium compound, and combining a FISH experiment to detect the interaction of TCAB1 with hTR;
b. searching a promoter sequence of a TCAB1 gene in a database, designing a primer for PCR amplification, carrying out 454 sequencing on a PCR product, and comparing the change of TCAB1 promoter sequences of a selenium compound positive group and a selenium compound negative group;
c. study of signaling pathway of selenium compounds to TCAB1 regulation: comparing the gene expression spectrum difference of selenium compound positive and selenium compound negative tumor cells by adopting a gene chip technology, carrying out clustering analysis and pathwa analysis by using bioinformatics GSEA (glutathione-acetyltransferase) to find out signal pathways participating in regulation of effector molecules (NF-kappa B, c-Myc, Sp1, PKC and p53), verifying the change of main signal molecules by using a real-time fluorescence quantitative PCR (polymerase chain reaction) or Western Blot technology, and determining a possible signal transduction pathway of selenium compound inactivation TCAB 1;
fishing of TCAB1 promoter binding protein: constructing TAP-TCAB1 adenovirus vector, infecting selenium compound positive and selenium compound negative groups, extracting total protein or nucleoprotein, using magnetic beads to specifically adsorb TCAB1-TAP and interaction protein complex, analyzing by MALDI-TOF, identifying polypeptide fingerprint, and screening signal protein (c-Myc, Sp1, PKC, p53) related to TCAB1 activation pathway.
Preferably, the specific research method in step three is as follows:
a. blocking the effect of the ATR pathway on TCAB1 expression and related signal molecules: the ATR pathway is identified according to the research results of a part of the steps, key signal molecules (ATR, H2AX, TopBP1, Chk1) in the pathway are blocked by ATR inhibitors or siRNA, and the expression of TCAB1, CB localization and related signal molecules (c-Myc, Sp1, PKC, p53) in cells of an active group and an inactive group of the selenium compounds are detected;
effect of tcab1 pathway gene silencing on ATR pathway: synthesizing TCAB1 siRNA according to the TCAB1 signal channel screened in the second step, detecting the DNA damage activation range under the action of selenium compound, and analyzing the expression of ATRIP and TopBP1 to detect the influence on the ATR channel.
Preferably, the specific research method in step three is as follows:
the method comprises the steps of treating selenium compound positive group tumor cells by using a genome stability intervention small molecule probe (ATR pathway inhibitor or TCAB1 pathway siRNA), observing indexes of proliferation, apoptosis and survival capability of aged cells of cells treated by combination camptothecin or radiation, and screening potential combined killing small molecule chemical substances.
(III) advantageous effects
The invention provides a molecular probe for treating head and neck squamous cell carcinoma and application thereof. The method has the following beneficial effects:
the invention further clarifies a molecular mechanism of activating ATR (attenuated total transcription) pathway signaling by a selenium compound by utilizing a molecular cytobiology technology, the molecular mechanism of reducing TCAB1 expression and signaling by the selenium compound, and the interaction of the two pathways and a mechanism causing instability of a genome and killing tumors, so that a small molecular probe of a genome stability intervention response pathway with an sensitization effect on cervical squamous cell carcinoma under the action of the selenium compound is screened, and a new thought and a new method are provided for the treatment of clinical head and neck squamous cell carcinoma.
Drawings
FIG. 1 is a flow chart of the research technique of the molecular probe for head and neck squamous carcinoma treatment of the present invention;
FIG. 2 is a schematic diagram of ATR pathway activation process of the molecular probe for head and neck squamous carcinoma therapy of the present invention;
FIG. 3 is a functional diagram of TCAB1 in telomere synthesis pathway of the molecular probe for treating head and neck squamous carcinoma according to the present invention;
FIG. 4 is a diagram of a cell culture according to the present invention;
FIG. 5 is a diagram of a colony formation experiment according to the present invention;
FIG. 6 is a Tunel assay of the present invention;
FIG. 7 is a diagram of fluorescent quantitative PCR according to the present invention;
FIG. 8 is a WB detection diagram according to the present invention;
FIG. 9 is a graph of co-immunoprecipitation according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
as shown in fig. 1 to 3, the embodiment of the present invention provides a molecular probe for head and neck squamous cell carcinoma therapy and an application thereof, and studies the influence of selenium compounds on the genome stability of head and neck squamous cell carcinoma cells:
researching the action of activating an ATR (attenuated total reflectance) channel by a selenium compound;
analyzing the action of the selenium compound on TCAB1 transcriptional inactivation;
step three, researching the interaction of the ATR channel and the TCAB1 channel;
analyzing the feasibility of the genome stability intervention treatment of the head and neck squamous cell carcinoma under the action of the selenium compound;
during research, the tumor cell genome includes two human oral squamous carcinoma cell lines (HSC-3, Cal-27), a nasopharyngeal carcinoma cell line CNE1 and an adenoid cystic carcinoma cell line ACC2, and the acting reagents of the selenium compound are sodium selenite and seleno-L-methionine.
The specific research method in the step one is as follows:
a. systemic analysis of molecular expression of ATR pathway signaling in selenium compound-treated head and neck squamous carcinoma cells: detecting the capability of forming foci of ATRIP, TopBP1, 9-1-1 and Mre11-Rad50-Nbs1 complex, H2AX, CTIP and PCNA signal molecules by immunofluorescence and immunoblotting;
b. formation of single-stranded DNA intermediate: detecting the forming capacity of single-stranded DNA (ssDNA) in the selenium compound processing head and neck squamous carcinoma cells by using a phosphorylation RPA and non-denaturation BrdU immunofluorescence method;
stability of dna replication forks: BrdU and IdU are used for labeling cells in an interactive mode, and the length of an IdU single-color fluorescence labeling area is observed after ultraviolet rays or camptothecin are used for treating head and neck squamous carcinoma cells so as to indicate the capacity of restarting and extending a replication fork;
d. screening of selenium compound acting protein: constructing an adenovirus vector, expressing selenium compound related protein with a TAP label, preparing a high-titer adenovirus vector, infecting head and neck squamous carcinoma cells (108 cell numbers), extracting total protein or nucleoprotein, specifically adsorbing a selenium compound TAP and an interaction protein complex by using magnetic beads (Dynal beads) which are covalently bonded with rabbit IgG, separating and cutting a specific protein band by SDS-PAGE electrophoresis, then carrying out MALDI-TOF or liquid chromatography-related mass spectrometry, identifying polypeptide fingerprints, and screening target protein related to an ATR (attenuated total reflectance) channel.
The specific research method in the step two is as follows:
a. influence of selenium compound on contents of components of telomerase complex and localization of Cajal body: TRAP-ELISA for detecting telomerase activity; detecting telomere length by Southern blot; westernblot detection of TCAB1, hTERT and dyskherin; notherblot detection of hTR levels; using antibodies of TCAB1, p80-coilin (a marker protein of CB), dyskerin and hTERT to carry out a co-immunoprecipitation experiment to detect the interaction of TCAB1 with dyskerin, hTERT and CB under the action of a selenium compound, and combining a FISH experiment to detect the interaction of TCAB1 with hTR;
b. searching a promoter sequence of a TCAB1 gene in a database, designing a primer for PCR amplification, carrying out 454 sequencing on a PCR product, and comparing the change of TCAB1 promoter sequences of a selenium compound positive group and a selenium compound negative group;
c. study of signaling pathway of selenium compounds to TCAB1 regulation: comparing the gene expression spectrum difference of selenium compound positive and selenium compound negative tumor cells by adopting a gene chip technology, carrying out clustering analysis and pathwa analysis by using bioinformatics GSEA (glutathione-acetyltransferase) to find out signal pathways participating in regulation of effector molecules (NF-kappa B, c-Myc, Sp1, PKC and p53), verifying the change of main signal molecules by using a real-time fluorescence quantitative PCR (polymerase chain reaction) or Western Blot technology, and determining a possible signal transduction pathway of selenium compound inactivation TCAB 1;
fishing of TCAB1 promoter binding protein: constructing TAP-TCAB1 adenovirus vector, infecting selenium compound positive and selenium compound negative groups, extracting total protein or nucleoprotein, using magnetic beads to specifically adsorb TCAB1-TAP and interaction protein complex, analyzing by MALDI-TOF, identifying polypeptide fingerprint, and screening signal protein (c-Myc, Sp1, PKC, p53) related to TCAB1 activation pathway.
The concrete research method in the third step is as follows:
a. blocking the effect of the ATR pathway on TCAB1 expression and related signal molecules: the ATR pathway is identified according to the research results of a part of the steps, key signal molecules (ATR, H2AX, TopBP1, Chk1) in the pathway are blocked by ATR inhibitors or siRNA, and the expression of TCAB1, CB localization and related signal molecules (c-Myc, Sp1, PKC, p53) in cells of an active group and an inactive group of the selenium compounds are detected;
effect of tcab1 pathway gene silencing on ATR pathway: synthesizing TCAB1 siRNA according to the TCAB1 signal channel screened in the second step, detecting the DNA damage activation range under the action of selenium compound, and analyzing the expression of ATRIP and TopBP1 to detect the influence on the ATR channel.
The concrete research method in the third step is as follows:
the method comprises the steps of treating selenium compound positive group tumor cells by using a genome stability intervention small molecule probe (ATR pathway inhibitor or TCAB1 pathway siRNA), observing indexes of proliferation, apoptosis and survival capability of aged cells of cells treated by combination camptothecin or radiation, and screening potential combined killing small molecule chemical substances.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (6)
1. The molecular probe for treating head and neck squamous carcinoma and the application thereof are characterized in that: the influence of selenium compounds on the stability of the genome of head and neck squamous cell carcinoma cells was studied:
researching the action of activating an ATR (attenuated total reflectance) channel by a selenium compound;
analyzing the action of the selenium compound on TCAB1 transcriptional inactivation;
step three, researching the interaction of the ATR channel and the TCAB1 channel;
analyzing the feasibility of the genome stability intervention treatment of the head and neck squamous cell carcinoma under the action of the selenium compound;
during research, the tumor cell genome comprises two human oral squamous carcinoma cell lines (HSC-3, Cal-27), a nasopharyngeal carcinoma cell line CNE1 and an adenoid cystic carcinoma cell line ACC2, and the acting reagents of the selenium compound are sodium selenite and seleno-L-methionine.
2. The molecular probe for head and neck squamous carcinoma therapy according to claim 1, characterized in that: the specific research method in the first step is as follows:
a. systemic analysis of molecular expression of ATR pathway signaling in selenium compound-treated head and neck squamous carcinoma cells: detecting the capability of forming foci of ATRIP, TopBP1, 9-1-1 and Mre11-Rad50-Nbs1 complex, H2AX, CTIP and PCNA signal molecules by immunofluorescence and immunoblotting;
b. formation of single-stranded DNA intermediate: detecting the forming capacity of single-stranded DNA (ssDNA) in the selenium compound processing head and neck squamous carcinoma cells by using a phosphorylation RPA and non-denaturation BrdU immunofluorescence method;
stability of dna replication forks: BrdU and IdU are used for labeling cells in an interactive mode, and the length of an IdU single-color fluorescence labeling area is observed after ultraviolet rays or camptothecin are used for treating head and neck squamous carcinoma cells so as to indicate the capacity of restarting and extending a replication fork;
d. screening of selenium compound acting protein: constructing an adenovirus vector, expressing selenium compound related protein with a TAP label, preparing a high-titer adenovirus vector, infecting head and neck squamous carcinoma cells (108 cell numbers), extracting total protein or nucleoprotein, specifically adsorbing a selenium compound TAP and an interaction protein complex by using magnetic beads (Dynal beads) which are covalently bonded with rabbit IgG, separating and cutting a specific protein band by SDS-PAGE electrophoresis, then carrying out MALDI-TOF or liquid chromatography-related mass spectrometry, identifying polypeptide fingerprints, and screening target protein related to an ATR (attenuated total reflectance) channel.
3. The molecular probe for head and neck squamous carcinoma therapy according to claim 1, characterized in that: the specific research method in the second step is as follows:
a. influence of selenium compound on contents of components of telomerase complex and localization of Cajal body: TRAP-ELISA for detecting telomerase activity; detecting telomere length by Southern blot; detecting TCAB1, hTERT and dyskherin by Western blot; notherblot detection of hTR levels; using antibodies of TCAB1, p80-coilin (a marker protein of CB), dyskerin and hTERT to carry out a co-immunoprecipitation experiment to detect the interaction of TCAB1 with dyskerin, hTERT and CB under the action of a selenium compound, and combining a FISH experiment to detect the interaction of TCAB1 with hTR;
b. searching a promoter sequence of a TCAB1 gene in a database, designing a primer for PCR amplification, carrying out 454 sequencing on a PCR product, and comparing the change of TCAB1 promoter sequences of a selenium compound positive group and a selenium compound negative group;
c. study of signaling pathway of selenium compounds to TCAB1 regulation: comparing the gene expression spectrum difference of selenium compound positive and selenium compound negative tumor cells by adopting a gene chip technology, carrying out clustering analysis and pathwa analysis by using bioinformatics GSEA (glutathione-acetyltransferase) to find out signal pathways participating in regulation of effector molecules (NF-kappa B, c-Myc, Sp1, PKC and p53), verifying the change of main signal molecules by using a real-time fluorescence quantitative PCR (polymerase chain reaction) or Western Blot technology, and determining a possible signal transduction pathway of selenium compound inactivation TCAB 1;
fishing of TCAB1 promoter binding protein: constructing TAP-TCAB1 adenovirus vector, infecting selenium compound positive and selenium compound negative groups, extracting total protein or nucleoprotein, using magnetic beads to specifically adsorb TCAB1-TAP and interaction protein complex, analyzing by MALDI-TOF, identifying polypeptide fingerprint, and screening signal protein (c-Myc, Sp1, PKC, p53) related to TCAB1 activation pathway.
4. The molecular probe for head and neck squamous carcinoma therapy according to claim 1, characterized in that: the concrete research method in the third step is as follows:
a. blocking the effect of the ATR pathway on TCAB1 expression and related signal molecules: the ATR pathway is identified according to the research results of a part of the steps, key signal molecules (ATR, H2AX, TopBP1, Chk1) in the pathway are blocked by ATR inhibitors or siRNA, and the expression of TCAB1, CB localization and related signal molecules (c-Myc, Sp1, PKC, p53) in cells of an active group and an inactive group of the selenium compounds are detected;
effect of tcab1 pathway gene silencing on ATR pathway: synthesizing TCAB1 siRNA according to the TCAB1 signal channel screened in the second step, detecting the DNA damage activation range under the action of selenium compound, and analyzing the expression of ATRIP and TopBP1 to detect the influence on the ATR channel.
5. The molecular probe for head and neck squamous carcinoma therapy according to claim 1, characterized in that: the concrete research method in the third step is as follows:
the method comprises the steps of treating selenium compound positive group tumor cells by using a genome stability intervention small molecule probe (ATR pathway inhibitor or TCAB1 pathway siRNA), observing indexes of proliferation, apoptosis and survival capability of aged cells of cells treated by combination camptothecin or radiation, and screening potential combined killing small molecule chemical substances.
6. Use of a molecular probe according to claims 1-5 for the treatment and inhibition of head and neck squamous carcinoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111325700.4A CN114015775A (en) | 2021-11-10 | 2021-11-10 | Molecular probe for treating head and neck squamous cell carcinoma and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111325700.4A CN114015775A (en) | 2021-11-10 | 2021-11-10 | Molecular probe for treating head and neck squamous cell carcinoma and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114015775A true CN114015775A (en) | 2022-02-08 |
Family
ID=80063207
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111325700.4A Pending CN114015775A (en) | 2021-11-10 | 2021-11-10 | Molecular probe for treating head and neck squamous cell carcinoma and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114015775A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090325201A1 (en) * | 2006-05-12 | 2009-12-31 | University Of Miami | Biomarkers for Detection and Diagnosis of Head and Neck Squamous Cell Carcinoma |
| US20140213472A1 (en) * | 2013-01-29 | 2014-07-31 | William Beaumont Hospital | Head and Neck Cancer Biomarkers |
| WO2018175673A1 (en) * | 2017-03-22 | 2018-09-27 | The Rockefeller University | Methods and compositions for detection and treatment of cancer |
| US20190330625A1 (en) * | 2016-06-14 | 2019-10-31 | Agency For Science, Technology And Research | Consequences of a defective switch in cutaneous squamous cell carcinoma |
-
2021
- 2021-11-10 CN CN202111325700.4A patent/CN114015775A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090325201A1 (en) * | 2006-05-12 | 2009-12-31 | University Of Miami | Biomarkers for Detection and Diagnosis of Head and Neck Squamous Cell Carcinoma |
| US20140213472A1 (en) * | 2013-01-29 | 2014-07-31 | William Beaumont Hospital | Head and Neck Cancer Biomarkers |
| US20190330625A1 (en) * | 2016-06-14 | 2019-10-31 | Agency For Science, Technology And Research | Consequences of a defective switch in cutaneous squamous cell carcinoma |
| WO2018175673A1 (en) * | 2017-03-22 | 2018-09-27 | The Rockefeller University | Methods and compositions for detection and treatment of cancer |
Non-Patent Citations (2)
| Title |
|---|
| CHONG-KUI SUN等: "TCAB1: a potential target fordiagnosis and therapy of head andneck carcinomas", 《MOLECULAR CANCER》 * |
| TINGTING QIN等: "Expressed HNSCC variants by HPV-status in a well-characterizedMichigan cohort", 《SCIENTIFIC REPORTS》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Huang et al. | Detection of Histone H3 mutations in cerebrospinal fluid-derived tumor DNA from children with diffuse midline glioma | |
| Zhang et al. | TP53 mutations in epithelial ovarian cancer | |
| Tomlins et al. | Integrative biology of prostate cancer progression | |
| Carles et al. | Head and neck squamous cell carcinoma transcriptome analysis by comprehensive validated differential display | |
| Rosell et al. | BRCA1: a novel prognostic factor in resected non-small-cell lung cancer | |
| Kawase et al. | PH domain-only protein PHLDA3 is a p53-regulated repressor of Akt | |
| Raetz et al. | Gene expression profiling reveals intrinsic differences between T‐cell acute lymphoblastic leukemia and T‐cell lymphoblastic lymphoma | |
| Langbein et al. | Metastasis is promoted by a bioenergetic switch: new targets for progressive renal cell cancer | |
| Bahl et al. | Novel polymorphism in p21waf1/cip1 cyclin dependent kinase inhibitor gene: association with human esophageal cancer | |
| Pan et al. | Integrated bioinformatics analysis of master regulators in anaplastic thyroid carcinoma | |
| Kawakami et al. | Detection of novel paraja ring finger 2‐fer tyrosine kinase mRNA chimeras is associated with poor postoperative prognosis in non‐small cell lung cancer | |
| Takita et al. | Gene expression profiling and identification of novel prognostic marker genes in neuroblastoma | |
| Lu et al. | O-GlcNAcylation: an important post-translational modification and a potential therapeutic target for cancer therapy | |
| Gao et al. | Gastric cardia adenocarcinoma microRNA profiling in Chinese patients | |
| Le Boiteux et al. | Widespread overexpression from the four DNA hypermethylated HOX clusters in aggressive (IDH wt) glioma is associated with H3K27me3 depletion and alternative promoter usage | |
| Shen et al. | Mitochondrial dysfunction pathway alterations offer potential biomarkers and therapeutic targets for ovarian cancer | |
| Marengo et al. | MYC expression and metabolic redox changes in cancer cells: a synergy able to induce chemoresistance | |
| Kapodistrias et al. | Assessing the clinical value of microRNAs in formalin-fixed paraffin-embedded liposarcoma tissues: Overexpressed miR-155 is an indicator of poor prognosis | |
| Toy et al. | Investigating molecular determinants of cancer cell resistance to ionizing radiation through an integrative bioinformatics approach | |
| Inokawa et al. | Detection of doublecortin domain-containing 2 (DCDC2), a new candidate tumor suppressor gene of hepatocellular carcinoma, by triple combination array analysis | |
| Chang et al. | Post-translational modifications of EMT transcriptional factors in cancer metastasis | |
| CN114015775A (en) | Molecular probe for treating head and neck squamous cell carcinoma and application thereof | |
| Garcia et al. | The GADD45, ZBRK1 and BRCA1 pathway: quantitative analysis of mRNA expression in colon carcinomas | |
| Cavalloni et al. | Proteomic analysis identifies deregulated metabolic and oxidative-associated proteins in Italian intrahepatic cholangiocarcinoma patients | |
| Jia et al. | IL24 and its receptors regulate growth and migration of pancreatic cancer cells and are potential biomarkers for IL24 molecular therapy |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230925 Address after: 445000 No. 2, Wufengshan Road, Tuqiao Avenue, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province Applicant after: Minda Hospital Affiliated to Hubei University for Nationalities Address before: 445000 No. 169, 4th lane, Wuyang Avenue, Enshi Tujia and Miao Autonomous Prefecture, Hubei Province Applicant before: Zhang Bo |
|
| TA01 | Transfer of patent application right |