CN-115478107-B - Gene composition, kit and detection method for detecting lung adenocarcinoma KRAS mutation

CN115478107BCN 115478107 BCN115478107 BCN 115478107BCN-115478107-B

Abstract

The invention relates to a gene composition, a kit and a method for detecting KRAS mutation of lung adenocarcinoma, belonging to the technical field of molecular diagnosis. The invention provides eighteen miRNA gene compositions for evaluating the KRAS mutation of lung adenocarcinoma for the first time, and compared with the current common method, the method has the advantages of convenient operation, quantitative analysis, high accuracy and good sensitivity and specificity.

Inventors

Bian Binyi
ZHAN CHENG
LIANG JIAQI
Sui Qihai
SHAN GUANGYAO
Hu zhengyang
FAN HONG
WANG QUN

Assignees

复旦大学附属中山医院

Dates

Publication Date: 20260505
Application Date: 20220927

Claims (7)

1. A genetic composition for detecting KRAS mutations in lung adenocarcinoma, comprising the following miRNA genes ：hsa-miR-141-3p、hsa-miR-1307-5p、has-miR-331-3p、hsa-miR-9-5p、hsa-miR-127-3p、hsa-miR-181c-5p、hsa-miR-125b-5p、hsa-miR-21-5p、hsa-miR-146b-5p、hsa-miR-197-3p、hsa-miR-146b-3p、hsa-miR-185-5p、hsa-miR-424-5p、hsa-miR-375-3p、hsa-miR-99a-5p、hsa-miR-21-3p、hsa-miR-374a-3p and has-let-7c-5p.
2. Use of the genetic composition of claim 1 for the manufacture of a medicament for detecting a KRAS mutation in lung adenocarcinoma.
3. The use according to claim 2, wherein the medicament comprises an agent for detecting the expression level of the gene composition and comprises the same as the sole active ingredient.
4. A kit for detecting KRAS mutation in lung adenocarcinoma, comprising a reagent for detecting the expression level of the gene composition of claim 1, wherein the reagent is used as the only active ingredient.
5. The kit for detecting a KRAS mutation in lung adenocarcinoma of claim 4, further comprising instructions for recording a threshold value and Score formula for the miRNA gene composition, wherein the threshold value for the miRNA gene composition is as follows: the Score scoring formula is as follows: Score=0.4360 hsa-miR-141-3p+0.4551 hsa-miR-1307-5p+0.2065 has-miR-331-3p+0.8180 hsa-miR-9-5p+0.3668 hsa-miR-127-3p+0.7692 hsa-miR-181c-5p+0.7516 hsa-miR-125b-5p+1.1773 hsa-miR-21-5p+0.5870 hsa-miR-146b-5p+0.8374 hsa-miR-197-3p+0.4296 hsa-miR-146b-3p+7.4558 hsa-miR-185-5p+9.7320 hsa-miR-424-5p+0.6608 hsa-miR-375-3p+8.1049 hsa-miR-99a-5p+0.7248 hsa-miR-21-3p+0.1435 hsa-miR-374a-3p+0.5374 has-let-7c-5p-21.6752。
6. the kit for detecting a KRAS mutation according to claim 5, wherein the detection sample is judged to have a KRAS mutation when the Score of the detection sample is higher than the cutoff value of the lung adenocarcinoma, and the detection sample is judged to have no KRAS mutation when the Score of the detection sample is not higher than the cutoff value of the lung adenocarcinoma, and wherein the cutoff value of the lung adenocarcinoma is-0.62.
7. The kit for detecting KRAS mutation in lung adenocarcinoma according to claim 6, wherein the detection sample is a fresh tissue lung adenocarcinoma sample.

Description

Gene composition, kit and detection method for detecting lung adenocarcinoma KRAS mutation Technical Field The invention belongs to the technical field of molecular diagnosis, and particularly relates to a gene composition, a kit and a method for detecting lung adenocarcinoma KRAS mutation. Background Lung cancer is the highest worldwide cancer with mortality rates in the first and second place in men and women, respectively, with an average five-year survival rate of less than 15%. Among them, lung adenocarcinoma is the main subtype of lung cancer at present, accounting for nearly 60% of new cases, and is characterized by low survival rate. The Kirsten murine sarcoma virus carcinoma (KIRSTEN RAT sarcoma viral oncogene, KRAS) gene mutation is one of the common mutations of lung adenocarcinoma, the mutation rate in lung cancer is 32% [1], playing a critical role in the development of tumors [1]. Point mutations are the most common way of KRAS mutation, the most common types being KRAS-G12D mutation (41%), KRAS-G12V mutation (28%) and KRAS-G12C mutation (14%). In NSCLC, G12C is the most common type [3], which is closely related to poor prognosis [4]. The targeted KRAS mutant medicine AMG510[5] has made great progress, sotorasib is marketed in 2021, and fills the blank of KRAS mutant targeted medicine. In view of the high risk of KRAS mutant lung adenocarcinoma, the KRAS mutant patient can be accurately identified early, and the method has extremely important significance for improving the targeted treatment effect of the patient and reducing the burden of society. The existing method for directly detecting KRAS mutation clinically has the problems of high cost, complicated flow, low accuracy and the like, and the detection method with higher sensitivity, specificity and application value can be applied to the establishment of an individual targeting treatment scheme of lung adenocarcinoma patients. Detecting the expression level of a plurality of miRNAs of a patient as a biomarker for judging the KRAS mutation condition of the patient can provide guarantee for KRAS mutation detection more accurately, and is beneficial to realizing accurate medical treatment for the patient. Disclosure of Invention In view of the shortcomings of the prior art, the invention provides a gene composition, a kit and a method for detecting lung adenocarcinoma KRAS mutation. In order to solve the problems, the invention adopts the following technical scheme: In a first aspect, the invention provides a gene composition for detecting a KRAS mutation in lung adenocarcinoma, comprising the following miRNA gene compositions ：hsa-miR-141-3p、hsa-miR-1307-5p、has-miR-331-3p、hsa-miR-9-5p、hsa-miR-127-3p、hsa-miR-181c-5p、hsa-miR-125b-5p、hsa-miR-21-5p、hsa-miR-146b-5p、hsa-miR-197-3p、hsa-miR-146b-3p、hsa-miR-185-5p、hsa-miR-424-5p、hsa-miR-375-3p、hsa-miR-99a-5p、hsa-miR-21-3p、hsa-miR-374a-3p and has-let-7c-5p. In a second aspect, the invention also provides an application of the gene composition in preparing a medicine for detecting lung adenocarcinoma KRAS mutation. Preferably, the drug comprises an agent for detecting the expression level of the gene composition, and uses it as the only active ingredient. In a third aspect, the present invention also provides a kit for detecting KRAS mutation in lung adenocarcinoma, comprising a reagent for detecting the expression level of the gene composition as described above, wherein the reagent is used as the only active ingredient. Preferably, the instructions further include instructions for recording a threshold value and Score formula for the miRNA gene, wherein the threshold value for the miRNA gene composition is as follows: the Score scoring formula is as follows: Score＝0.4360*hsa-miR-141-3p+0.4551*hsa-miR-1307-5p+0.2065*has-miR-331-3p+0.8180*hsa-miR-9-5p+0.3668*hsa-miR-127-3p+0.7692*hsa-miR-181c-5p+0.7516*hsa-miR-125b-5p+1.1773*hsa-miR-21-5p+0.5870*hsa-miR-146b-5p+0.8374*hsa-miR-197-3p+0.4296*hsa-miR-146b-3p+7.4558*hsa-miR-185-5p+9.7320*hsa-miR-424-5p+0.6608*hsa-miR-375-3p+8.1049*hsa-miR-99a-5p+0.7248*hsa-miR-21-3p+0.1435*hsa-miR-374a-3p+0.5374*has-let-7c-5p-21.6752. Preferably, the detection sample is judged to have KRAS mutation when the Score of the detection sample is higher than the cutoff value of lung adenocarcinoma, and the detection sample is judged to have no KRAS mutation when the Score of the detection sample is not higher than the cutoff value of lung adenocarcinoma, wherein the cutoff value of lung adenocarcinoma is-0.62. Preferably, the test sample is a fresh tissue lung adenocarcinoma sample. In a fourth aspect, the present invention also provides a method for detecting KRAS mutation of lung adenocarcinoma, comprising the steps of: Step (1), collecting a detection sample, and detecting the expression level ：hsa-miR-141-3p、hsa-miR-1307-5p、has-miR-331-3p、hsa-miR-9-5p、hsa-miR-127-3p、hsa-miR-181c-5p、hsa-miR-125b-5p、hsa-miR-21-5p、hsa-miR-146b-5p、hsa-miR-197-3p、hsa-miR-146b-3p、hsa-miR-185-5p、h