CN108646020A - Purposes of the Girdin albumen in preparing diagnosis of pancreatic cancer preparation and treatment of pancreatic cancer drug - Google Patents

Purposes of the Girdin albumen in preparing diagnosis of pancreatic cancer preparation and treatment of pancreatic cancer drug Download PDF

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CN108646020A
CN108646020A CN201810494250.3A CN201810494250A CN108646020A CN 108646020 A CN108646020 A CN 108646020A CN 201810494250 A CN201810494250 A CN 201810494250A CN 108646020 A CN108646020 A CN 108646020A
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leu
glu
lys
girdin
ser
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王芸
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    • 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
    • G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups

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Abstract

The present invention provides a kind of purposes of Girdin albumen in preparing diagnosis of pancreatic cancer preparation, and the amino acid sequence of the Girdin albumen is as shown in SEQ NO.1.The present invention also provides a kind of purposes of Girdin albumen in preparing treatment of pancreatic cancer drug, and the amino acid sequence of the Girdin albumen is as shown in SEQ NO.1.The present invention has the following technical effect that：(1) since for Girdin protein expressions higher than cancer beside organism, the expression for passing through Girdin albumen and its relevant components ingredient in the individual pancreatic tissue of detection and (or) blood increases the generation for predicting cancer of pancreas in Pancreas cancer patients cancerous tissue；(2) since Girdin albumen is to the facilitation of pancreatic cancer growth, cancer of pancreas is treated by reducing the expression of Girdin albumen and its relevant components ingredient in Pancreatic Adenocarcinoma；(3) since Girdin takes part in the relevant pathologic process of the cellular activities such as malignant proliferation, apoptosis, invasion, the present invention provides experience and application foundation to the relationship of further investigated Girdin and other diseases from now on.

Description

Girdin albumen is in preparing diagnosis of pancreatic cancer preparation and treatment of pancreatic cancer drug Purposes
Technical field
The present invention provides the purposes of Girdin albumen, belong to biomedicine technical field.
Background technology
Cancer of pancreas is that a kind of grade malignancy is very high, all highly difficult malignant tumor of digestive tract of diagnosing and treating, 5 years survival rates ＜ 1% is worst one of the malignant tumour of prognosis.Its morbidity and mortality obviously rises in recent years, occupies mortality of malignant tumors The 4th of reason, incidence of the annual death rate nearly equivalent to current year.The diagnosis rate of cancer of pancreas early stage is not high, and operation is dead It is higher to die rate, and cure rate is very low.The treatment of cancer of pancreas at present mainly based on surgical intervention, supplemented by radiotherapy, chemotherapy, and Inquire into the new method for combining the biological therapies such as immune and molecule.The high reason of the death rate of cancer of pancreas mainly when finding It is in middle and advanced stage and gradually transfer.Based on this, clinically must actively improve the early diagnosis of cancer of pancreas, treatment level and Improve the prognosis of patient.
Girdin is a kind of new actin binding protein, is a macromolecular being made of 1870 amino acid residues Protein, and be named as Akt phosphorylation enhancer, with cell to migrate movement closely related.In newborn nerve cell and swell Girdin expression quantity can be significantly raised in oncocyte, is generated with new vessels also closely related.There is experiment to find the height of Girdin Expression is closely related with the progress of cancer of pancreas, and Girdin genes participate in the occurrence and development of cancer of pancreas.Since Giridn albumen can swash PIK3/Akt signal transducting systems living, and the activation of PI3K/Akt signal paths participates in the generation of cancer of pancreas, Girdin can lead to Cross the occurrence and development process of PI3/AKt signal paths regulation and control cancer of pancreas.Girdin is played during the occurrence and development of cancer of pancreas Important function, this prompt further can be used for antitumor control using Girdin as diagnosis of pancreatic cancer and as target spot It treats.
The domestic and foreign literature and patent of the prior art are retrieved, so far there are no any Girdin albumen and its relevant group At ingredient for the research of pancreatic cell diagnosis of pancreatic cancer and treatment report.
Invention content
The purpose of the present invention is to overcome the deficiencies of the prior art, provides a kind of Girdin albumen and examines preparing cancer of pancreas Purposes in disconnected preparation, the amino acid sequence of the Girdin albumen is as shown in SEQ NO.1.
It is a further object to provide a kind of purposes of Girdin albumen in preparing treatment of pancreatic cancer drug, institutes The amino acid sequence for the Girdin albumen stated is as shown in SEQ NO.1.
The present invention has the following technical effect that：
(1) Girdin protein expressions are higher than cancer beside organism in Pancreas cancer patients cancerous tissue, by detecting individual pancreatic tissue And (or) the expression of Girdin albumen and its relevant components ingredient increases to predict the generation of cancer of pancreas in blood；
(2) Girdin albumen has facilitation to pancreatic cancer growth, by reducing Girdin albumen in Pancreatic Adenocarcinoma And its expression of relevant components ingredient treats cancer of pancreas；
(3) since Girdin takes part in the relevant pathologic process of the cellular activities such as malignant proliferation, apoptosis, invasion, this Invention provides experience and application foundation to the relationship of further investigated Girdin and other diseases from now on.
Description of the drawings
Figure 1A is cancer of pancreas Girdin protein immunizations histochemical staining (× 100) result；
Figure 1B is cancer beside organism Girdin protein immunizations histochemical staining (× 100) result；
Fig. 2 is that the shRNA retrovirus of Girdin knocks out the result of Girdin albumen；
The shRNA retrovirus that Fig. 3 A are Girdin knocks out Girdin and inhibits the transfer of cancer of pancreas PANC-1 cell invasions Cell scratch experiment result；
The shRNA retrovirus that Fig. 3 B are Girdin knocks out Girdin and inhibits the transfer of cancer of pancreas PANC-1 cell invasions Cell migration assay result；
The shRNA retrovirus that Fig. 3 C are Girdin knocks out Girdin and inhibits the transfer of cancer of pancreas PANC-1 cell invasions Cell invasion experimental result；
The shRNA retrovirus that Fig. 4 is Girdin lowers the result that Girdin inhibits cancer of pancreas PANC-1 cell Proliferations；
The shRNA retrovirus that Fig. 5 is Girdin lowers the result that Girdin promotes cancer of pancreas PANC-1 Apoptosis；
The shRNA retrovirus that Fig. 6 is Girdin lowers the influence that Girdin grows tumor bearing nude mice Pancreatic Adenocarcinoma As a result.
Specific implementation mode
It is further illustrated the present invention below in conjunction with specific embodiment.It is noted that following embodiment is served only for illustrating The present invention, rather than limit the scope of the invention.
The Girdin albumen mentioned in the following example is amino acid sequence Girdin albumen as shown in SEQ NO.1.
Embodiment 1
Girdin protein expressions are higher than the clinical meaning of cancer beside organism in Pancreas cancer patients cancerous tissue
Using immunohistochemistry Envision two step methods detection Girdin albumen by by 90 cancers of pancreas and 90 cancers The expression in the organization chip of composition is organized, and combines clinical and pathological data for statistical analysis.As a result as Figure 1A -1B, Shown in table 1,2, show that Girdin protein expressions positive rate is higher than cancer beside organism (P in Pancreatic Adenocarcinoma<0.05), but with it is sexual Not, the not statistically significant (P of correlation of age, tumor size, the clinical stages of cancer of pancreas, pathological grading etc.>0.05).
These results prompt expression of the Girdin albumen in cancer of pancreas to increase, may take part in the generation of cancer of pancreas with into Exhibition, reference frame is provided for the diagnosis and Index for diagnosis of cancer of pancreas.
Expression of the table 1Girdin in cancer of pancreas and cancer beside organism
The relationship of table 2Girdin protein expressions and cancer of pancreas clinical pathologic characteristic
Embodiment 2
Confirm Girdin in pancreas carcinogenesis and the effect in progress in cell model level
(1) the shRNA retrovirus of transfection Girdin inhibits the growth of pancreatic cancer cell and promotes pancreatic cancer cell Apoptosis
As shown in Fig. 2, with the shRNA retrovirus of Girdin in pancreatic cancer cell PANC-1, passing through Western Blot detection Girdin protein expression levels verification retrovirus knocks out effect, as a result shows in pancreatic cancer cell PANC-1 Girdin protein expressions are obviously lowered, and show that this Girdin high-expression vectors can effectively knock out Girdin.
It is transfected using retrovirus in pancreatic cancer cell PANC-1, it is real by cell cut, migration, invasion It tests, it was confirmed that the knockout of Girdin makes the transfer of cell slow down, it is suppressed that the invasion of cancer cell are shifted, and have statistical significance (* * P ＜ 0.01, as shown in figs 3 a and 3b).After the shRNA retrovirus for transfecting Girdin 48 hours, detected by CCK-8 Cell proliferative conditions, the results showed that at 48 hours, this Girdin knocks out identical as the speed of growth of cellular control unit system.However 96 hours, the PANC-1 cell growths after Girdin is knocked out were decreased obviously (as shown in Figure 4).It is thin in order to further study cancer of pancreas Girdin biological meanings in born of the same parents, Apoptosis detection are compared using APC/PI dyeing, and compared with the control group, Girdin is knocked out Cell in apoptosis rate obviously increase (as shown in Figure 5).The expression downward of result above prompt Girdin albumen inhibits pancreas The growth of cancer cell and the apoptosis for promoting pancreatic cancer cell.
Embodiment 3 confirms inhibiting effect of the Girdin albumen to pancreatic cancer growth in animal model level
Using the cancer of pancreas kind tumor experiment for the cell, cellular control unit progress nude mice for confirming Girdin knockouts, by PANC- 1 groups of cells, PANC-1+Girdin gene upregulations and downward groups of cells cell are incubated respectively, are centrifuged, and dilution is planted in nude mice armpit Under nest, observation plantation latter week, two weeks, three weeks, the growing state of surrounding knurl respectively.It is put to death respectively by the above-mentioned period naked Mouse takes out knurl, finds the PANC-1 cell seedings after knocking out Girdin with nude mice, the speed of tumor formation obviously subtracts Slowly, tumor formation rate is low, and on the nude mice of normal PANC-1 tumor formations, after the shRNA retrovirus for injecting Giridn, knurl is apparent Become smaller.Its size between control group and direct cell knockout group nude mice (as shown in Figure 6), the experimental results showed that Girdin albumen Downward can inhibit the growth of Pancreatic Adenocarcinoma in tumor bearing nude mice.
In conclusion the expression raising of Girdin albumen plays an important role in the occurrence and development of pancreatic cell cancer of pancreas, It can be increased come pre- by detecting the expression of Girdin albumen and its relevant components ingredient in individual pancreatic tissue and (or) blood The generation for surveying cancer of pancreas, by the expression for reducing Girdin albumen and its relevant components ingredient in individual and (or) Pancreatic Adenocarcinoma Level is used for the treatment of cancer of pancreas, Girdin albumen and its diagnosis, Index for diagnosis and life that relevant components ingredient is cancer of pancreas Object treatment provides new reference frame.
SEQUENCE LISTING
<110>Wang Yun
<120>Purposes of the Girdin albumen in preparing diagnosis of pancreatic cancer preparation and treatment of pancreatic cancer drug
<160> 1
<170> PatentIn version 3.3
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<211> 1870
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Ser Arg Gln Ser Lys Asp Ser Asn Pro Tyr Ala Thr Leu Pro Arg
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Val Asp Ser Pro Pro Ala Ala Ala Asp Ser Asn Thr Thr Ala Ala
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Claims (2)

1. a kind of purposes of Girdin albumen in preparing diagnosis of pancreatic cancer preparation, which is characterized in that the Girdin albumen Amino acid sequence as shown in SEQ NO.1.
2. a kind of purposes of Girdin albumen in preparing treatment of pancreatic cancer drug, which is characterized in that the Girdin albumen Amino acid sequence as shown in SEQ NO.1.
CN201810494250.3A 2018-05-22 2018-05-22 Purposes of the Girdin albumen in preparing diagnosis of pancreatic cancer preparation and treatment of pancreatic cancer drug Pending CN108646020A (en)

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