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Human paclitaxel-resistant gastric cancer cells hgc-27/ptx and preparation method thereof

A technology of HGC-27, gastric cancer cells, applied in the field of biomedicine, can solve the problems such as hindering the continued benefit of patients, and achieve the effect of stable traits

Active Publication Date: 2019-06-25
BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the prolongation of medication time, paclitaxel resistance seriously hinders patients from continuing to benefit

Method used

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  • Human paclitaxel-resistant gastric cancer cells hgc-27/ptx and preparation method thereof
  • Human paclitaxel-resistant gastric cancer cells hgc-27/ptx and preparation method thereof
  • Human paclitaxel-resistant gastric cancer cells hgc-27/ptx and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0041] The preparation method of the CCK-8 working solution is as follows: 10 volume parts of CCK-8 and 90 volume parts of RPMI-1640 complete medium are mixed. CCK-8 is a product of Tongren Chemical Research Institute in Japan.

[0042] Relative activity calculation formula:

[0043]

[0044] The formula for calculating drug resistance index:

[0045] Drug resistance index = half inhibitory concentration of HGC-27 / PTX / half inhibitory concentration of HGC-27 / DZ.

Embodiment 1

[0046] Example 1. Establishment of Human Paclitaxel-resistant Gastric Cancer Cells

[0047] 1. Establishment of human paclitaxel-resistant gastric cancer cells

[0048] Human paclitaxel-resistant gastric cancer cells were constructed by gradually increasing the concentration of paclitaxel and intermittently induced in vitro. The specific steps are as follows:

[0049] 1. Preparation of HGC-27 suspension

[0050] Human gastric cancer cell line HGC-27 was trypsinized with 0.25% EDTA, and then "RPMI-1640 complete medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin" was added , placed at 37°C, 5% CO 2 cultured in an incubator to obtain a cell density of 72×10 5 individual / mL HGC-27 suspension.

[0051] 2. Preparation of culture system

[0052] Take the cell culture dish, add RPMI-1640 complete medium, HGC-27 suspension and paclitaxel injection to obtain a culture system. In this culture system, the concentration of human gastric cancer cell l...

Embodiment 2

[0066] Example 2, the sensitivity of HGC-27 / PTX to paclitaxel, cisplatin and fluorouracil

[0067] 1. The sensitivity of HGC-27 / PTX to paclitaxel

[0068] Set up the experimental group, and then perform the following steps:

[0069] (1) Cultivate the cells to be tested (HGC-27 / PTX or HGC-27 / DZ) to obtain a cell suspension.

[0070] (2) Take a 96-well plate and inoculate each well with 100 μL of the cell suspension prepared in step (1) (each well contains about 1×10 cells to be tested). 4 ), placed at 37°C, 5% CO 2 Cultivate in the incubator for 24h.

[0071] (3) After completing step (2), take the 96-well plate and add paclitaxel injection to obtain a treatment system; when the cells to be tested are HGC-27 / PTX, the concentration of paclitaxel in the treatment system is 2.5nM, 5nM , 10nM, 50nM, 250nM or 1250nM (3 replicate wells are set for each paclitaxel concentration); 32nM, 64nM or 128nM (three replicate wells were set for each paclitaxel concentration).

[0072] (...

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Abstract

The invention discloses a human gastric cancer taxol drug resistance cell HGC-27 / PTX and a preparation method of the human gastric cancer taxol drug resistance cell HGC-27 / PTX. The human gastric cancer taxol drug resistance cell HGC-27 / PTX is preserved in the China General Microbiological Culture Collection Center (CGMCC), and is assigned with the accession number of CGMCC No.13596. The human gastric cancer taxol drug resistance cell HGC-27 / PTX provided by the invention is stable in property, and can realize a plurality of times of passage stably, so that the in-depth study can be carried out on a molecular mechanism causing cellular drug resistance, and the human gastric cancer taxol drug resistance cell HGC-27 / PTX is an ideal model for studying the drug resistance mechanism of the patient suffering from gastric cancer, and has a significant application value.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to human paclitaxel-resistant gastric cancer cells HGC-27 / PTX and a preparation method thereof. Background technique [0002] Gastric cancer is the third leading cause of cancer death worldwide, and its morbidity and mortality both rank second among malignant tumors in my country. In my country, more than 70% of patients are already in the advanced stage or advanced stage when they are diagnosed, and they lose the opportunity for radical surgery. Drug therapy is the main method for advanced gastric cancer. Among the three types of chemotherapy drugs for gastric cancer, paclitaxel is widely used in the treatment of gastric cancer due to its high efficacy and good tolerance. However, with the prolongation of medication time, paclitaxel resistance seriously hinders patients from continuing to benefit. The key to solving the problem is to study the mechanism of paclitaxel resistance and exp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2501/06C12N2503/02G01N33/5011G01N2500/10
Inventor 沈琳高静李艳艳
Owner BEIJING CANCER HOSPITAL PEKING UNIV CANCER HOSPITAL
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