CN114908167A - Application of SVIL expression level detection reagent in preparation of ovarian cancer dryness diagnosis reagent - Google Patents
Application of SVIL expression level detection reagent in preparation of ovarian cancer dryness diagnosis reagent Download PDFInfo
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- CN114908167A CN114908167A CN202210477305.6A CN202210477305A CN114908167A CN 114908167 A CN114908167 A CN 114908167A CN 202210477305 A CN202210477305 A CN 202210477305A CN 114908167 A CN114908167 A CN 114908167A
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Abstract
本发明公开了SVIL表达水平检测试剂在制备卵巢癌干性诊断试剂中的应用；SVIL基因全称为Supervillin，其氨基酸包含细胞核定位功能域，羧基端包含肌动蛋白结合功能域，功能与招募细胞骨架蛋白至细胞膜和细胞核中的特定位置相关。SVIL表达水平与卵巢癌干性相关，因此，SVIL表达水平的检测试剂可应用于卵巢癌干性诊断试剂的制备。本发明可用于评价卵巢癌干性，可用于为卵巢癌恶性程度和患者生存期提供参考。
The invention discloses the application of SVIL expression level detection reagents in the preparation of ovarian cancer stemness diagnostic reagents; the full name of SVIL gene is Supervillin, its amino acid contains a nuclear localization functional domain, its carboxyl terminus contains an actin binding functional domain, and its function is related to recruiting cytoskeleton. Proteins are associated to specific locations in the cell membrane and nucleus. The expression level of SVIL is related to the stemness of ovarian cancer. Therefore, the detection reagent for the expression level of SVIL can be applied to the preparation of the diagnostic reagent for the stemness of ovarian cancer. The invention can be used to evaluate the stemness of ovarian cancer, and can be used to provide reference for the malignant degree of ovarian cancer and the survival period of patients.
Description
Technical Field
The invention relates to the field of medical molecular test diagnosis, in particular to an application of an SVIL expression level detection reagent in preparation of an ovarian cancer dryness diagnosis reagent.
Background
Ovarian cancer is a great threat to female life, the recurrence rate of ovarian cancer is as high as 70%, the five-year survival rate is less than 40%, and the ovarian cancer is one of the most malignant tumors. Ovarian cancer stem cells are one of the major causes of high malignancy of ovarian cancer. The ovarian cancer stem cells are cells with stem cell functions such as self-renewal in tumor tissues, and have stronger tumor forming capability, metastatic invasion capability, drug resistance capability and the like compared with differentiated ovarian cancer stem cells, so that identification of ovarian cancer sternness is an important reference for making treatment strategies and evaluating prognosis of patients, but no effective ovarian cancer sternness evaluation marker and corresponding detection method exist at present.
The SVIL gene is collectively referred to as Supervillin, and its amino acids comprise a nuclear localization domain and its carboxy-terminus comprises an actin binding domain, functions related to recruiting cytoskeletal proteins to specific locations in the cell membrane and nucleus. In the research of the applicant, the expression level of the SVIL is in inverse proportion to the dryness of the ovarian cancer and in direct proportion to the survival time of the patient, and the SVIL is suggested to have the effect of inhibiting the dryness of the ovarian cancer stem cells. Therefore, the detection of the expression level of SVIL can be used for judging the dryness of ovarian cancer and further judging the survival time of the patient. Therefore, the invention provides a new method for judging the dryness of the ovarian cancer, which can be used for judging the malignancy of the ovarian cancer and the survival prognosis of patients.
Disclosure of Invention
The invention aims to provide an application of a SVIL expression level detection reagent in preparation of an ovarian cancer dryness diagnosis reagent.
In order to achieve the purpose, the invention provides the following technical scheme:
application of SVIL expression level detection reagent in preparation of ovarian cancer dryness diagnosis reagent.
Preferably, the SVIL is called supervillin, the ID in Pubmed data is 6840, and the position in human Chromosome is Chromosome 10, NC-000010.11 (29457338 and 29736936).
Preferably, the expression level of SVIL refers to the mRNA expression level and the protein expression level of SVIL in the cell.
In a preferred embodiment of the invention, the method for detecting the expression of SVILmRNA comprises a probe method, a qRT-PCR method and a sequencing method.
The invention is preferable, and the method adopted by the reagent for detecting the SVIL protein expression comprises western blot, immunohistochemistry and mass spectrometry.
Preferably, the dryness of the ovarian cancer refers to the dryness of ovarian cancer cells on an orthotopic site, a metastasis, ascites and peritoneum, and also includes the dryness of primary cultured ovarian cancer cells and ovarian cancer cell line cells.
According to the invention, the meaning of clinical detection of the dryness of the ovarian cancer is that the high dryness of the ovarian cancer cells represents high malignancy of the ovarian cancer cells.
Preferably, the ovarian cancer dryness fraction has clinical diagnostic significance that the ovarian cancer dryness fraction is high and represents the average life cycle of a corresponding patient population is short.
Preferably, the ovarian cancer dryness is characterized in vitro by being proportional to the ovarian cancer cell dryness and the microsphere forming capacity of the ovarian cancer cells.
The invention has the beneficial effects that: the invention discloses the application of a reagent for detecting the expression level of SVIL (singular value decomposition) in preparing an ovarian cancer dry diagnostic reagent for the first time. The method for detecting the SVIL mRNA expression level reagent comprises a probe method, a qRT-PCR method and a sequencing method. The method for detecting the SVIL protein expression level reagent comprises western blot, immunohistochemistry and mass spectrum. The reagent can detect the dryness of ovarian cancer cells in situ, metastasis, ascites and peritoneum, and can also detect the dryness of primary cultured ovarian cancer cells and ovarian cancer cell line cells. The invention provides a new way for detecting the dryness degree of ovarian cancer.
Drawings
In order to make the purpose, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a graph showing that SVIL expression in TCGA database ovarian cancer tissue samples is proportional to patient survival;
FIG. 2 shows the accuracy of the expression level of SVIL in determining the dryness of ovarian cancer cells cultured in vitro.
Detailed Description
The present invention is further described below in conjunction with the following figures and specific examples so that those skilled in the art may better understand the present invention and practice it, and the experimental methods are not described in the examples, but generally according to the conventional conditions or according to the conditions recommended by the manufacturers. The examples are not intended to limit the invention.
Example 1 expression of SVIL in ovarian cancer tissue samples from the TCGA database in proportion to patient survival
The mRNA and miRNA expression data of the ovarian cancer patient are downloaded from a TCGA (the cancer genome atlas) database, the expression data of SVIL and the follow-up data of the patient are extracted from the database, after the data are washed and processed, the patient is divided into a high expression group and a low expression group according to the expression level of SVIL, the Kaplan-Meier method is used for analyzing the correlation between the expression of SVIL and the long-term survival of the patient, and the result is shown in figure 1, and the low expression of SVIL is related to the poor prognosis of the patient (P is less than 0.05).
Example 2 accuracy of determination of dryness of ovarian cancer cells cultured in vitro by expression level of SVIL
The method comprises the following steps of culturing the cells of ovarian cancer, such as Caov-3, SKOV3, A2780, Ovcar4, ES2, Ovcar8, Caov-4, Nihovcar3, Heya8 and IGROV1, separately culturing the microspheres of ovarian cancer cells in a suspension culture mode, culturing adherent ovarian cancer cells in a traditional adherent culture mode, and respectively detecting the RNA level of SVIL of each cell, wherein the primers used in RT-PCR detection are as follows:
SVILF：5'-gacacccctcgatacatgaga-3'(SEQ ID NO.1)；
SVIL R：5'-cggaggtttctgtgcagtatt-3'(SEQ ID NO.2)；
GAPDH F：5’-ctgggctacactgagcacc-3’(SEQ ID NO.3)；
GAPDH R：5’-aagtggtcgttgagggcaatg-3’(SEQ ID NO.4)。
using formula CT (SVIL-GAPDH) >7 judging the cells with high dryness degree by using the formula CT (SVIL-GAPDH) Judging the cells with low dryness degree less than or equal to 7, and culturing the cells in suspensionThe spheres are cells with high actual dryness, adherent ovarian cancer cells cultured by a traditional adherent culture mode are cells with low actual dryness, and the accuracy of the dryness of the ovarian cancer cells cultured in vitro is judged by analyzing the SVIL expression level through an ROC curve. As a result, the AUC value obtained by determining the dryness of ovarian cancer cells cultured in vitro based on the expression level of SVIL was 0.7978, as shown in FIG. 2. The result indicates that the method has better accuracy.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitutions or changes made by the person skilled in the art on the basis of the present invention are all within the protection scope of the present invention. The protection scope of the invention is subject to the claims.
Sequence listing
<110> Chongqing university affiliated tumor hospital
Application of <120> SVIL expression level detection reagent in preparation of ovarian cancer dry diagnostic reagent
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
gacacccctc gatacatgag a 21
<210> 2
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
cggaggtttc tgtgcagtat t 21
<210> 3
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
ctgggctaca ctgagcacc 19
<210> 4
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
aagtggtcgt tgagggcaat g 21
Claims (9)
- Application of a SVIL expression level detection reagent in preparation of ovarian cancer dryness diagnosis reagents.
- 2. Use according to claim 1, characterized in that: the SVIL is known as supervillin, ID in Pubmed data is 6840, and position in human Chromosome is Chromosome 10, NC-000010.11 (29457338 and 29736936).
- 3. Use according to claim 1, characterized in that: the expression level of SVIL refers to the mRNA expression level and the protein expression level of SVIL in the cell.
- 4. Use according to claim 1, characterized in that: the method adopted by the reagent for detecting the expression of the SVILmRNA comprises a probe method, a qRT-PCR method and a sequencing method.
- 5. Use according to claim 1, characterized in that: the method adopted by the reagent for detecting the SVIL protein expression comprises western blot, immunohistochemistry and mass spectrometry.
- 6. Use according to claim 1, characterized in that: the dryness of the ovarian cancer refers to the dryness of ovarian cancer cells on an orthotopic area, a metastasis area, ascites and peritoneum, and also comprises the dryness of primary cultured ovarian cancer cells and ovarian cancer cell line cells.
- 7. Use according to claim 1, characterized in that: the clinical detection significance of the ovarian cancer dryness is that the high dryness degree of ovarian cancer cells represents the high malignancy degree of ovarian cancer cells.
- 8. Use according to claim 1, characterized in that: the clinical diagnosis significance of the ovarian cancer dryness is that the ovarian cancer dryness is high and represents the average life cycle of corresponding patient groups.
- 9. Use according to claim 1, characterized in that: the ovarian cancer dryness is characterized in that the dryness of ovarian cancer cells is proportional to the microsphere forming capacity of the ovarian cancer cells.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1922304A (en) * | 2003-10-31 | 2007-02-28 | 维特克公司 | Blood test prototype and method for detecting circulating tumor and endothelial cells |
| CN109475596A (en) * | 2016-03-08 | 2019-03-15 | 伊玛提克斯生物技术有限公司 | Uterine Cancer Treatment |
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- 2022-05-04 CN CN202210477305.6A patent/CN114908167A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1922304A (en) * | 2003-10-31 | 2007-02-28 | 维特克公司 | Blood test prototype and method for detecting circulating tumor and endothelial cells |
| CN109475596A (en) * | 2016-03-08 | 2019-03-15 | 伊玛提克斯生物技术有限公司 | Uterine Cancer Treatment |
Non-Patent Citations (1)
| Title |
|---|
| WON-TAE KIM 等: "Cancer stem cell surface markers on normal stem cells", BMB REP, vol. 50, no. 6, 31 December 2017 (2017-12-31), pages 285 - 298 * |
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