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Kit for detecting drug resistance of malignant parenchymatous tumors to chemotherapeutic drugs

A detection kit and technology for solid tumors, applied in the direction of microbial determination/inspection, biochemical equipment and methods, etc., can solve problems such as increasing drug efflux

Active Publication Date: 2016-11-16
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the level of tumor cells, there are different mechanisms of drug resistance, including increasing drug efflux, changing drug targets, repairing DNA damage, activating other pathways, etc., but how to overcome the drug resistance of tumor cells has always been an open question. unsolved problems

Method used

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  • Kit for detecting drug resistance of malignant parenchymatous tumors to chemotherapeutic drugs
  • Kit for detecting drug resistance of malignant parenchymatous tumors to chemotherapeutic drugs
  • Kit for detecting drug resistance of malignant parenchymatous tumors to chemotherapeutic drugs

Examples

Experimental program
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Effect test

Embodiment 1

[0054] Verification of the expression and correlation analysis of MDR1 and LIMK1 in drug-resistant malignant solid tumor cells

[0055] In order to verify the expression and characteristics of MDR1 and LIMK1 in malignant solid tumor drug-resistant cells, the present invention established differential drug-resistant cell sublines by intermittently exposing malignant solid tumor cells to vincristine (VCR) with a gradient increase. The drug resistance index was calculated and graded according to the drug resistance test. Collect drug-resistant cell samples from each group, and collect sensitive parental cell samples as a control group, and use fluorescent real-time quantitative PCR (qRT-PCR) for verification. The specific steps are as follows:

[0056] (1), RNA extraction

[0057] ①Take the tumor cells cultured to the logarithmic growth phase and wash them 3 times with pre-cooled PBS. Add 1ml Trizol to each bottle to lyse the cells and shake horizontally on ice for 5min. Use a...

Embodiment 2

[0074] Verification of the expression and correlation analysis of MDR1 and LIMK1 in drug-resistant malignant solid tumor tissues

[0075] In the present invention, drug-resistant cells of low-, medium- and high-grade malignant solid tumors that have been successfully constructed are respectively inoculated into nude mice to establish a nude mouse drug-resistant model. Collect resistant tissue samples from each group, use non-drug-resistant tissue samples as controls, and use fluorescent real-time quantitative PCR (qRT-PCR) for verification. The specific operation steps are as follows:

[0076] (1), RNA extraction

[0077] ①Collect the tumor tissue and freeze it in liquid nitrogen. After taking it out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered, add Trizol and leave it at room temperature for 5 minutes;

[0078] ② Add 0.2ml of 1 / 5 volume of chloroform, vibrate the centrifuge tube vigorously, mix well, and place it at room temperat...

Embodiment 3

[0095] Preparation of Drug Resistance Diagnostic Kit for Malignant Solid Tumors

[0096] According to the relationship between MDR1 and LIMK1 genes and drug resistance in malignant solid tumors, as well as the law of expression changes in drug-resistant cells (tissues), by simultaneously detecting the expression of MDR1 gene and LIMK1 gene, and calculating the ratio of the two gene expression ratios, it is possible to judge the Whether or not the patient is drug-resistant and the degree of drug resistance plays an important guiding role in the precise selection of individualized drugs for tumor chemotherapy regimens.

[0097] Further, the chemotherapeutic drugs involved in the present invention are limited to conventional clinical chemotherapeutic drugs such as doxorubicin, paclitaxel, pirarubicin, methotrexate, and gemcitabine.

[0098] Further, the malignant solid tumors involved in the present invention are limited to lung cancer, gastric cancer, liver cancer, colon cancer,...

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Abstract

The invention discloses a kit for detecting drug resistance of malignant parenchymatous tumors to chemotherapeutic drugs. The kit comprises malignant parenchymatous tumor drug-resistant cell cDNA standard samples, an SYBR Green polymerase chain reaction system, MDR1 and LIMK1 genes for amplification and housekeeping gene primer pairs. When the kit is used, cells of the malignant parenchymatous tumors of a lung cancer and the like are intermittently exposed in vincristine with increasing gradients, and a differential drug-resistant cell substrain is established. It is detected and found through a fluorescent quantitative PCR instrument that expression of the MDR1 gene is enhanced along with increasing of the drug-resistant degree, and expression of the LIMK1 gene is enhanced in cells which have low and moderate resistance to the drugs and retraced in cells which have high resistance to the drugs. Therefore, whether the malignant parenchymatous tumors are resistant to the drugs or not and the drug-resistant degree of the malignant parenchymatous tumors can be diagnosed by jointly detecting expressions of the MDR1 and LIMK1 genes; meanwhile, detection of the expressions of the MDR1 and LIMK1 genes can be used for preparing a chemotherapeutic drug resistance screening preparation, and an accurate therapeutic regimen can be provided for a patient subjected to malignant parenchymatous tumor chemotherapy.

Description

technical field [0001] The invention relates to a detection kit for chemotherapeutic drug resistance of malignant solid tumors Background technique [0002] Since the 1970s, the Ministry of Health and other departments have conducted many surveys on the cause of death across the country, and the results show that the mortality rate of malignant tumors in my country is increasing year by year. From 1973 to 1975, the total malignant tumor mortality rate of men and women in my country was 74.2 / 100,000; from 1990 to 1992, this figure rose to 108.26 / 100,000, an increase of 45.9%; from 2004 to 2005, it had climbed to 135.88 / 100,000, which has nearly doubled in 30 years. By 2010, it rose to 162.87 / 100,000. Among them, the increase in the morbidity and mortality rate of lung cancer is the primary reason for the increase in the total mortality rate of malignant tumors, followed by colon, rectum, anal cancer, and breast cancer. [0003] Chemotherapy combined with surgery has always...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 王岩
Owner JILIN UNIV
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