Human stomach cancer multidrug-resistant cell strain

A multidrug resistance and cell line technology for gastric cancer, applied in the field of cell engineering

Inactive Publication Date: 2014-04-23
JIANGSU KEYGEN BIOTECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Through searching, it is found that there is no literature report on the establishment of human gastric cancer multi-drug resistant cell line SGC-7901 / VP by using the human gastric cancer cell line SGC-7901 as the induction object at present.

Method used

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  • Human stomach cancer multidrug-resistant cell strain
  • Human stomach cancer multidrug-resistant cell strain
  • Human stomach cancer multidrug-resistant cell strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Induction and establishment of human gastric cancer multidrug-resistant cell line SGC-7901 / VP

[0021] The human gastric cancer multidrug-resistant cell line SGC-7901 / VP was established by the in vitro induction method of gradually increasing VP concentration and intermittent action. The specific steps are as follows:

[0022] (1) Culture medium for human gastric cancer cell line SGC-7901 (RPMI-1640 medium containing 10% calf serum), 5% CO2, 37°C, culture to 70%-90% adherence rate, remove Add VP100ng / ml impact culture for 1 hour to the supernatant, remove the VP100ng / ml culture solution, wash the blank RPMI-1640 culture solution twice, replace the proliferation medium and culture for 72-96h to expand the surviving cells. When the surviving cells amplify to 70%-90% adherence rate, repeat the impact culture with VP100ng / ml culture solution for 1 hour and 9 times, that is, complete 10 times of 100ng / ml VP culture solution impact culture for 1 hour in total, and f...

Embodiment 2

[0025] Morphological observation of embodiment 2 drug-resistant strains

[0026] Take the human gastric cancer multidrug-resistant cell line SGC-7901 / VP in the logarithmic growth phase, and observe the cell morphology under an inverted microscope. The results are as follows: Figure 6 Shown: the cell shape becomes larger, the nucleus shrinks, and the cells are easy to accumulate into clusters.

Embodiment 3

[0027] Example 3 Cell Growth Curve Determination

[0028] Cells were digested, counted, and prepared into a cell suspension with a concentration of 5×103 / mL, and 100 μL of cell suspension was added to each well of a 96-well cell culture plate (500 cells per well); the 6-well cell culture plate was placed at 37°C , cultured in a 5% CO2 incubator for 2d, 4d, 6d, 8d, and 10d respectively; stain the 96-well plate with MTT, λ=490nm, and measure the OD value; add 20μL MTT (5mg / mL) to each well, and continue in the incubator Incubate for 4 hours; discard the medium, add 150 μL DMSO to each well to dissolve, shake gently for 10 minutes; λ=490nm, read the OD value of each well with a microplate reader, take time as the abscissa, and OD value as the ordinate Coordinates to plot cell growth curves. For cell growth curves of SGC-7901 and SGC-7901 / VP, see figure 1 .

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Abstract

The invention discloses a human stomach cancer multidrug-resistant cell strain which is established by employing human stomach cancer cell strain SGC-7091 as an induction object, employing a large-dose impact method and then employing an in-vitro induction method with gradually increased VP concentration and intermittent action. The cell strain is preserved China General Microbiology Center of Committee for Culture Collection of Microorganisms at 14th November, 2013, and the preservation number is CGMCC NO. 8455. The cell strain provided by the invention has typical multidrug-resistant characteristic, and provides an experiment base for further researching drug-resistant reversion approach.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and relates to a human cell strain, in particular to a human gastric cancer multidrug-resistant cell strain and a method for establishing the same. Background technique [0002] Malignant tumor is a serious disease that endangers people's health and life. In many countries and regions, it has accounted for the first or second cause of death among residents. Gastric cancer accounts for a considerable proportion of all malignant tumors. Its incidence rate and case fatality rate rank fourth and second among malignant tumors, respectively. In my country, the incidence and mortality of gastric cancer rank first in malignant tumors, and there is an upward trend. [0003] Chemotherapy is one of the important means to treat human malignant tumors. One of the reasons for the poor effect of chemotherapy is the generation of tumor drug resistance, and the reason has not been fully elucidated. Ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12R1/91
Inventor 不公告发明人
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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