CN114921548A - Application of ZNF526 in preparing liver cancer diagnosis and/or prognosis and treatment preparation and diagnosis, prognosis and treatment preparation - Google Patents
Application of ZNF526 in preparing liver cancer diagnosis and/or prognosis and treatment preparation and diagnosis, prognosis and treatment preparation Download PDFInfo
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Abstract
The invention discloses an application of ZNF526 in preparing a liver cancer diagnosis and/or prognosis and treatment preparation and the diagnosis, prognosis and treatment preparation. The invention discovers that ZNF526 is closely related to the diagnosis and prognosis of liver cancer for the first time and is a key gene for tumorigenesis and development, and patients with low expression of ZNF526 gene have higher survival rate. Therefore, compared with the prior art, the new scheme for diagnosing and treating the liver cancer by taking the ZNF526 gene as the molecular marker has stronger clinical application value and potential.
Description
Technical Field
The invention belongs to the technical field of medical molecular biology, and particularly relates to application of ZNF526 gene in preparation of liver cancer diagnosis and/or prognosis and treatment preparations, and diagnosis, prognosis and treatment preparations.
Background
Primary liver cancer is one of the common malignant tumors, and the mortality rate of primary liver cancer ranks third in all tumors, seriously threatening human health. China is one of high-incidence areas of liver cancer, and the number of the incidence of the liver cancer accounts for more than 50 percent of the whole world. Liver cancer has the characteristics of rapid progress, poor prognosis, short survival period and the like, and the five-year survival rate is less than 20%. Liver cancer is heterogeneous, has obvious difference in prognosis, but has no effective molecular marker for prognosis evaluation in clinic and has no target gene and related medicine for accurate treatment. The invention identifies the target gene ZNF526 closely related to the prognosis of the liver cancer, has the potential of becoming a target for subtype diagnosis, drug screening and treatment of the disease, provides theoretical and practical basis for clinically and effectively treating the liver cancer, and fills up the technical blank of target treatment of the liver cancer in international and domestic.
Disclosure of Invention
The invention aims to provide a molecular marker for effectively diagnosing liver cancer clinically, provide a molecular target for clinically searching a medicament and a strategy for effectively treating liver cancer and further achieve the overall prognosis effect of the liver cancer.
The invention provides an application of a ZNF526 gene detection reagent in preparing liver cancer diagnosis and/or prognosis preparations, wherein the CDS sequence of the ZNF526 gene is shown as SEQ ID NO. 1. ZNF526 zinc finger protein 526, Homo sapiens (human), Gene ID:116115
Further, the liver cancer diagnosis and/or prognosis preparation comprises a reagent for detecting the ZNF526 gene expression quantity by PCR or in situ hybridization.
Furthermore, the reagent for PCR detection of ZNF526 gene expression level comprises the following primer sequences:
ZNF 526-F: 5'-CGGGATCCATGGCAGAGGTGGTGGCTGAGGTG-3', respectively; as shown in SEQ ID NO. 2;
ZNF 526-R: 5'-CGACGCGTGCACGAAGGCCGTGTCCAACT-3', as shown in SEQ ID NO. 3.
The invention provides a liver cancer diagnosis and/or prognosis preparation, which comprises a reagent for detecting ZNF526 gene expression quantity by PCR.
Further, the liver cancer diagnosis and/or prognosis preparation comprises the primer.
The invention provides an application of a reagent for inhibiting ZNF526 gene expression in preparing a preparation for treating liver cancer.
Further, the agent for inhibiting ZNF526 gene expression comprises: shRNA.
The shRNA sequence is as follows:
shRNA1-ZNF526-F：5'-CCGGACTTTGACTCCCTAGAGAAAGCTCGAGCTT
TCTCTAGGGAGTCAAAGTTTTTTG-3'; as shown in SEQ ID NO. 4;
shRNA1-ZNF526-R：5'-AATTCAAAAAACTTTGACTCCCTAGAGAAAGCTC
GAGCTTTCTCTAGGGAGTCAAAGT-3'; as shown in SEQ ID NO. 5;
or:
shRNA2-ZNF526-F:5'-CCGGCCTTGCCTCATCCCTCTTCTTCTCGAGAAG
AAGAGGGATGAGGCAAGGTTTTTG-3'; as shown in SEQ ID NO. 6;
shRNA2-ZNF526-R:5'-AATTCAAAAACCTTGCCTCATCCCTCTTCTTCTCG
AGAAGAAGAGGGATGAGGCAAGG-3'; shown as SEQ ID NO.7
The invention also provides a preparation for treating liver cancer, which comprises an agent for inhibiting ZNF526 gene expression.
Further, the reagent for inhibiting ZNF526 gene expression comprises shRNA.
The invention relates to a tumor subtype classification, drug screening and treatment strategy taking a molecular marker ZNF526(zinc finger protein 526) as a target point. The ZNF526 is a target molecule for improving the overall prognosis of the liver cancer and effectively treating diseases; the ZNF526 gene is taken as a core target spot to intervene the expression of the gene, and is a feasible scheme for effectively treating liver cancer.
The invention has the technical advantages that:
ZNF526 is closely related to the prognosis of liver cancer, is a key gene for tumorigenesis and development, and has higher survival rate for patients with low expression of ZNF526 gene. Therefore, compared with the prior art, the new scheme for diagnosing and treating the liver cancer by taking the ZNF526 gene as the molecular marker has stronger clinical application value and potential.
The invention is further illustrated by the following description in conjunction with the accompanying drawings and detailed description, without limiting the invention thereto.
Drawings
FIG. 1 shows the mRNA expression level of ZNF526 in clinical liver cancer tissues and normal tissues; (P <0.0001, P <0.001, data of statistical significance)
FIG. 2 shows the overall survival rate (OS) and disease-free survival rate (DFS) of patients with high and low expression of ZNF526 of liver cancer patients;
fig. 3A cell viability of HepG2, Hep3B cells overexpressing ZNF526 and control (results mean ± standard deviation (n ═ 3); P <0.001,; P <0.05, data with statistical differences); HepG2 is a human hepatoblastoma cell line and Hep3B is a human hepatoma cell line, both purchased from the Shanghai cell Bank of the Chinese academy of sciences.
Fig. 3B knockdown cell viability for Huh-7, PVTT cells and control group of ZNF526 (mean ± standard deviation (n ═ 3); + P <0.05, data with statistical difference); huh-7 is a human liver cancer cell line, and PVTT is a human liver cancer thrombus cell line which are purchased from Shanghai cell banks of Chinese academy of sciences.
FIG. 4A: a.20 times of under-lens knockdown of cell migration conditions of HepG2 and Hep3B cells of ZNF526 and a control group; b. counting with Image J (results are mean ± standard deviation (n ═ 3); P <0.0001,; P <0.001, data with statistical differences);
FIG. 4B: cell migration of Huh-7 and PVTT cells over-expressing ZNF526 under a 20-fold mirror and a control group; b. counting with Image J (mean ± standard deviation (n ═ 3); (0.01); (0.001); (statistical differences in data);
FIG. 5: subcutaneous tumorigenesis of HepG2 nude mice a. tumors formed after subcutaneous injection of HepG2 cells (OEZNF526) and Control cells (Control) nude mice; b. tumors formed after subcutaneous injection of HepG2 cells (shRNA1-ZNF526) and control cells (Scramble) nude mice were knocked down (P <0.01, data are statistically different).
Detailed Description
Example 1: qPCR detection of ZNF526 expression in liver cancer tumor tissue and normal tissue
The liver cancer samples are all from hepatobiliary surgery of Xiangya II hospital, the samples are collected with consent of patients, and the samples are approved by the ethical committee of the institute of Life sciences. The fresh liver cancer sample and the matched normal liver tissue obtained by the surgical operation are quickly put into liquid nitrogen and then put into an ultra-low temperature refrigerator at minus 80 ℃ for storage. Smashing the liver cancer and normal tissues into powder by using a mortar at low temperature, adding 1mL of RZ lysate solution on ice for dissolving, transferring into a 1.5mL centrifuge tube, adding 200 mu L of trichloromethane, repeatedly reversing and uniformly mixing, standing on ice for 8min, and centrifuging at 14000rpm and 4 ℃ for 18 min. Transferring 400. mu.L of supernatant into a new 1.5mL centrifuge tube, adding 400. mu.L of isopropanol, mixing, placing on ice for 10min, and centrifuging at 14000rpm and 4 ℃ for 18 min. After the solution is discarded, the solution is washed for 2 times by 75 percent ethanol, is instantaneously centrifuged at 4 ℃, is discarded, is placed at room temperature until the ethanol is completely volatilized, is added with a proper amount of RNase-free water for dissolving, and is reversely transcribed into cDNA for detecting the gene transcription level.
The ZNF526 primer sequence used in the qPCR process was:
ZNF526-F：5'-CGGGATCCATGGCAGAGGTGGTGGCTGAGGTG-3'；
ZNF526-R：5'-CGACGCGTGCACGAAGGCCGTGTCCAACT-3'。
the experimental results are as follows:
FIG. 1 shows that for 8 pairs of clinical liver cancer tissues and normal tissues, the mRNA expression level of ZNF526 is higher in the liver cancer tumor tissues than in the normal tissues as shown by the qPCR result. (P <0.0001, P <0.001, data of statistical significance)
Example 2: prognostic assay for ZNF526
Among liver cancer patients, the prognosis of ZNF526 high-expression patients is poorer, and the overall survival period and the disease recurrence-free survival period are shorter than those of low-expression patients (figure 2).
The Overall survival rate (OS) and Disease free survival rate (DFS) of 181 liver cancer patients are analyzed, a Kaplan-Meier curve is drawn, the Overall survival time (P0.042) and Disease free survival time (P0.00097) of ZNF526 high-expression patients are obviously shorter than those of ZNF526 low-expression patients, the fact that the ZNF526 high-expression is related to poor prognosis of liver cancer is shown, and the fact that ZNF526 may play the role of a protooncogene in the occurrence and development of the liver cancer is suggested.
Example 3
CCK8 experiments show that ZNF526 overexpression can promote liver cancer cell proliferation (FIG. 3A), and ZNF526 knock-down can inhibit liver cancer cell proliferation (FIG. 3B).
Control refers to an over-expression empty vector plasmid;
OE-ZNF526 refers to ZNF526 over-expression plasmid;
scramble refers to knock-down of empty vector plasmid;
shRNA1-ZNF526 refers to ZNF526 knock-down sequence 1 plasmid;
shRNA2-ZNF526 refers to ZNF526 knock-down sequence 2 plasmid;
the method comprises the following steps:
a. an overexpression vector: pHAGE-full-EF1a-MCS-IZsGreen (purchased from Hainan Jia and Biotechnology Limited liability Co., Ltd.), with a size of 8246 bp;
enzyme cutting site: BamHI: 5'-C ^ GATCC-3'/XbaI: 5'-T ^ CTAGA-3'
And carrying out double enzyme digestion and connection on the ZNF526 gene fragment and a plasmid vector, and obtaining the over-expression ZNF526 plasmid after correct sequencing.
b. Knocking down the vector: pLKO.1 (purchased from Kyowa and Biotechnology, Inc.), 7032bp in size
Enzyme cutting site: age I: 5'-A ^ CCGGT-3'/EcoR I: 5'-G ^ AATTC-3'
Two short hairpin structure shRNAs are designed,
shRNA1-ZNF526-F：5'-CCGGACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGTTTTTTG-3'；
shRNA1-ZNF526-R：5'-AATTCAAAAAACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGT-3'；
shRNA2-ZNF526-F：5'-CCGGCCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGGTTTTTG-3'；
shRNA2-ZNF526-R：5'-AATTCAAAAACCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGG-3'。
and (3) annealing the shRNA primer, connecting the shRNA primer with the plasmid vector subjected to double enzyme digestion, and sequencing correctly to obtain the knockdown ZNF526 plasmid. Cell proliferation assay: CCK-8 detection
1) Taking cells in logarithmic phase, adding 0.5mL of pancreatin into each hole of a six-hole plate, shaking uniformly, placing in an incubator at 37 ℃ for standing and digestion, adding 2mL of complete culture medium into the holes to stop digestion after complete digestion, and blowing to homogenize cell suspension;
2) taking a clean blood counting chamber and a cover glass, counting cells, diluting the cells with a complete culture medium into cell suspension containing 3 ten thousand cells per mL, adding 100 mu L of cell suspension into each hole of a 96-hole plate to ensure that the number of the cells in each hole is 3000, arranging three multiple holes in each group, and respectively culturing the cells in a cell culture box with the temperature of 37 ℃ and the concentration of 5% CO2 for 24 hours, 48 hours, 72 hours and 96 hours;
3) diluting a CCK-8 reagent by using a cell growth culture medium according to the proportion of 1:10 to prepare a CCK-8 reagent culture medium, sucking and removing supernatant of a 96-well plate by using a suction filter, adding 100 mu L of prepared CCK-8 solution into each well at the same time point on different days, and incubating for 4 hours in a cell incubator at 37 ℃;
4) and measuring the light absorption value at 450nm of the microplate reader.
Example 4
Transwell experiments show that the ZNF526 is over-expressed to promote the migration of the liver cancer cells (FIG. 4A), and the ZNF526 is knocked down to inhibit the migration of the liver cancer cells (FIG. 4B). The method comprises the following steps:
cell migration assay: transwell experiment
1) Preparing a clean Transwell chamber in advance, soaking in 75% alcohol overnight, ventilating on a sterile operating table, irradiating with ultraviolet for 30min, and air drying;
2) to cells grown in log phase in six well plates, 0.5mL of pancreatin was added per well at 37 ℃ with 5% CO 2 Digesting in a cell incubator, adding 2mL of serum-free culture medium to stop digesting, taking a clean blood counting plate and a cover glass for counting cells, and diluting the cells into cell suspension containing 3 ten thousand cells per mL by using the culture medium containing no serum;
3) adding 600 mu L of complete culture medium containing 10% serum into each hole of a clean 24-hole plate, sleeving a Transwell chamber on each hole by using forceps, uniformly blowing cell suspension diluted by the serum-free culture medium, sucking 200 mu L of suspension liquid, hanging and dripping the suspension liquid in the center of the chamber, and culturing the suspension liquid in a 5% CO2 cell culture box at 37 ℃ for 48 hours;
4) taking a clean 24-pore plate, adding 600 mu L of 4% paraformaldehyde into each pore, slightly cleaning the small chamber in PBS by using a pair of forceps, sleeving the small chamber on the pore plate filled with the paraformaldehyde, and fixing for 30 min;
5) taking a clean 24-pore plate, adding 600 mu L of crystal violet solution into each pore, slightly washing the small chamber by PBS, sleeving the small chamber with the crystal violet, dyeing for 20min, washing away the loose color of the small chamber by PBS, wiping water drops in the small chamber by a cotton swab, and observing and counting by a microscope.
Example 5
The nude mouse tumorigenicity experiment shows that the tumorigenicity ability of the ZNF526 over-expressing tumor cells is increased (FIG. 5a), and the tumorigenicity ability of the ZNF526 knocking down the tumor cells is weakened (FIG. 5 b). The method comprises the following steps:
the experimental animals are subjected to examination by animal ethics committee of the university of south China, 8 BALB/c nude mice of 4 weeks are purchased from animal institute of south China to Spanish Sciaka laboratory animals Limited liability company of Hunan, and are maintained in an SPF level experimental barrier environment of animal institute of south China, and the animals are raised in an independent ventilated cage for one week after arrival. Meanwhile, HepG2ZNF526 overexpression, knockdown and control cells are cultured by a large dish, and the cells are uniformly digested respectively for cell counting, so that the cell density is 15 multiplied by 10 7 1mL total, 1mL matrigel was added and immediately transferred to ice after mixing to prevent the matrigel from setting. Before subcutaneous injection, the cells and the matrigel are mixed uniformly again by a pipette, a 1mL insulin needle is used for sucking cell suspension, the needle is slowly inserted into the two sides of the back of the forelimb of a nude mouse, subcutaneous injection is carried out, the cells of an experimental group HepG2 (OE-ZNF526 or shRNA1-ZNF526) are shot on the right side, the cells of a HepG2 Control group (Control or Scramble) are shot on the left side, the injection volume is 200 mu L on each side, and the cell number is 3 multiplied by 10 6 And forming a white round swelling bag under the skin of the mouse, slightly staying for a moment when withdrawing the needle, and carefully drawing the needle to prevent the cell suspension from overflowing and forming a strip-shaped tail. After the subcutaneous injection, the animals are continuously raised in the barrier environment of the department of zoology to observe the formation of tumors. Sacrifice mice using cervical dislocation, peel off subcutaneous tumors with clean scissors and forceps, record the photograph, measure the volume with a vernier caliper, then store inIn a formaldehyde solution.
Sequence listing
<110> university of Zhongnan
Application of <120> ZNF526 in preparation of liver cancer diagnosis and/or prognosis and treatment preparation and diagnosis, prognosis and treatment preparation
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2013
<212> DNA
<213> Intelligent (Homo sapiens)
<400> 1
atggcagagg tggtggctga ggtggccgag atgccaacac agatgtcacc aggggcagtg 60
gagatgtcaa cacctatgtc ggcagagatg atggagatgt caacagaagt gactgagatg 120
acacctgggg aggcccttgc ctcatccctc ttcttccagc atcaccagtt catgtgctct 180
gagtgtggca gcctctataa cacactggag gaagtcctct cacaccagga gcagcacatg 240
cttgctgtct cagaggagga ggcactgacc acacagaatg ttggcctgga gccggagctg 300
gtgccgggtg ctgaggggcc cttccagtgt ggtgaatgca gccagctcat cctctcccct 360
ggggagctcc tggcccacca ggatgcccac ctccgagagt ctgcaaacca gatccaatac 420
cagtgctggg actgccagga gctgttcccc tcgcccgagc tgtgggtggc tcatcgaaag 480
gcccagcacc tttctgctac ggtagctgag ccaccagtgc cacctccttt gcctccccca 540
acaccactgc ctccaccttc tcccccatcc gaagtcaaga tggagcccta tgagtgtcct 600
gagtgctcta ccctctgcgc cacccctgag gagttcttgg agcatcaggg cacccacttt 660
gactccctag agaaagagga gcgcaatggg ttggaggagg aggaagagga cgatgaggag 720
gatgaagaag atgatgaaga gatggaggat gaggaggcca tggcagaggt cggtgatgat 780
gctgtgggag gtgacgagtc cacagctggc tgggctcagg gctgcgggga ctgtccccag 840
caccagccct cagcaggggc tcgccggcaa caccggcgga cggctcacag cccggcatct 900
gccacccacc ccttccactg cagccagtgt cagcgcagtt tcagctccgc caaccggctg 960
caggctcatg ggcgggccca tgttggtggc acacatgagt gtacaacctg ctccaaggtc 1020
ttcaagaaag cagcatcgct tgagcagcac ttgcggctgc atcgcgggga agcccgctac 1080
ctctgtgtag actgtggccg cggctttggc acagaactca cgttggtggc tcaccggcgg 1140
gcccacactg ccaacccatt gcatcgctgt cgttgcggca agacgttcag caacatgacc 1200
aagttcctct accaccggcg cactcacgcc ggcaaaagcg gggcacctcc cacaggagca 1260
acagctcccc cagctccagc ggagcccacc cctccaccac caccccctgc cccacctgcc 1320
cagctgccct gcccacagtg ctccaagtcc tttgcctcag cttcccggct gtcccggcac 1380
cggcgtgcag tacacgggcc ccctgaacgg cgtcaccgct gtggggtttg tggcaagggc 1440
ttcaagaagc tgatccacgt gcgcaaccac ctgcggacac acacgggtga gaggcccttc 1500
cagtgccact catgtggcaa gacctttgct tctttggcca acctcagccg ccaccagctg 1560
acccatacgg gtgcacgtcc ctaccaatgc ctggactgtg gcaagcgctt cacacagagc 1620
tccaacctgc agcagcaccg gcggttgcac ttgcggccag tcgcctttgc ccgcgccccc 1680
cgcctcccca tcactggtct ctacaacaag agtccctact actgcgggac ttgtggccgc 1740
tggttccgcg ccatggcggg cttgcgactg catcagcggg tccatgcccg agctcggact 1800
ttgacgctac agcctcccag atcaccatct cctgccccac ccccacctcc agagcctcaa 1860
cagactatca tgtgcacaga gctgggggag accatcgcca tcattgagac atcccagcca 1920
ctggcgcttg aggacaccct gcagctgtgc caggctgcac tgggggccag tgaagcaggc 1980
gggctcttgc agttggacac ggccttcgtg tga 2013
<210> 2
<211> 32
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
cgggatccat ggcagaggtg gtggctgagg tg 32
<210> 3
<211> 29
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
cgacgcgtgc acgaaggccg tgtccaact 29
<210> 4
<211> 58
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
ccggactttg actccctaga gaaagctcga gctttctcta gggagtcaaa gttttttg 58
<210> 5
<211> 58
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
aattcaaaaa actttgactc cctagagaaa gctcgagctt tctctaggga gtcaaagt 58
<210> 6
<211> 58
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
ccggccttgc ctcatccctc ttcttctcga gaagaagagg gatgaggcaa ggtttttg 58
<210> 7
<211> 58
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
aattcaaaaa ccttgcctca tccctcttct tctcgagaag aagagggatg aggcaagg 58
Claims (10)
1. Application of a reagent for detecting ZNF526 gene in preparation of liver cancer diagnosis and/or prognosis preparations.
2. The use as claimed in claim 1, wherein the liver cancer diagnosis and/or prognosis agent comprises a reagent for detecting the expression level of ZNF526 gene by PCR or in situ hybridization.
3. The use as claimed in claim 1, wherein the reagent for PCR detection of ZNF526 gene expression level comprises the following primer sequences:
ZNF526-F：5'-CGGGATCCATGGCAGAGGTGGTGGCTGAGGTG-3'；
ZNF526-R：5'-CGACGCGTGCACGAAGGCCGTGTCCAACT-3'。
4. a liver cancer diagnosis and/or prognosis preparation is characterized by comprising a reagent for detecting the ZNF526 gene expression quantity by PCR.
5. The liver cancer diagnostic and/or prognostic preparation according to claim 4, comprising the primer set forth in claim 3.
6. The application of the reagent for inhibiting ZNF526 gene expression in preparing a preparation for treating liver cancer.
7. The use as claimed in claim 6 wherein the agent which inhibits expression of ZNF526 gene comprises: shRNA.
8. The use according to claim 7, wherein the shRNA has the following sequence:
shRNA1-ZNF526-F：5'-CCGGACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGTTTTTTG-3'；
shRNA1-ZNF526-R：5'-AATTCAAAAAACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGT-3'；
or:
shRNA2-ZNF526-F:5'-CCGGCCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGGTTTTTG-3'；
shRNA2-ZNF526-R:5'-AATTCAAAAACCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGG-3'。
9. a preparation for treating liver cancer, which is characterized by comprising an agent for inhibiting ZNF526 gene expression.
10. The agent for treating liver cancer according to claim 9, wherein the agent shRNA for inhibiting ZNF526 gene expression; the following sequences are preferred:
shRNA1-ZNF526-F：5'-CCGGACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGTTTTTTG-3'；
shRNA1-ZNF526-R：5'-AATTCAAAAAACTTTGACTCCCTAGAGAAAGCTCGAGCTTTCTCTAGGGAGTCAAAGT-3'；
or alternatively
shRNA2-ZNF526-F：5'-CCGGCCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGGTTTTTG-3'；
shRNA2-ZNF526-R：5'-AATTCAAAAACCTTGCCTCATCCCTCTTCTTCTCGAGAAGAAGAGGGATGAGGCAAGG-3'。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011106709A2 (en) * | 2010-02-26 | 2011-09-01 | Board Of Regents, The University Of Texas System | Epithelial biomarkers for cancer prognosis |
| WO2019018441A1 (en) * | 2017-07-17 | 2019-01-24 | Massachusetts Institute Of Technology | Cell atlas of healthy and diseased barrier tissues |
| CN113874505A (en) * | 2019-07-10 | 2021-12-31 | 核酸制药公司 | Aptamer specifically combined with KRAS protein and using method thereof |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011106709A2 (en) * | 2010-02-26 | 2011-09-01 | Board Of Regents, The University Of Texas System | Epithelial biomarkers for cancer prognosis |
| WO2019018441A1 (en) * | 2017-07-17 | 2019-01-24 | Massachusetts Institute Of Technology | Cell atlas of healthy and diseased barrier tissues |
| CN113874505A (en) * | 2019-07-10 | 2021-12-31 | 核酸制药公司 | Aptamer specifically combined with KRAS protein and using method thereof |
Non-Patent Citations (2)
| Title |
|---|
| XINXIN LI等: "Structures and biological functions of zinc finger proteins and their roles in hepatocellular carcinoma", 《BIOMARKER RESEARCH》, vol. 10, no. 2, 9 January 2022 (2022-01-09), pages 1 - 13 * |
| 周承汇;李年丰;: "ZNF207在肝细胞癌中的表达及意义", 中南大学学报(医学版), no. 04, 15 April 2019 (2019-04-15), pages 68 - 74 * |
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