Human choriocarcinoma multidrug resistant cell lines

A technology of drug-resistant cells and cell lines, applied in tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problem of choriocarcinoma patients not being able to achieve the purpose of curing, and achieve the effect of good drug resistance stability

Active Publication Date: 2019-06-28
ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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Problems solved by technology

[0003] However, there are still about 20% of chorioca...

Method used

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  • Human choriocarcinoma multidrug resistant cell lines
  • Human choriocarcinoma multidrug resistant cell lines
  • Human choriocarcinoma multidrug resistant cell lines

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Experimental program
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Embodiment Construction

[0016] The drug-resistant cell line establishment steps of the present invention are as follows:

[0017] (1) Human choriocarcinoma JAR cell line, after resuscitation, use RPMI-1640 medium (containing 10% fetal bovine serum) for routine culture in 5% CO2, 37 ° C incubator; human choriocarcinoma JEG3 cell line, after resuscitation, use EBSS-MEM medium (containing 10% fetal bovine serum) was routinely cultured in an incubator with 5% CO2 and 37°C;

[0018] (2) Take the JAR and JEG3 cells in the logarithmic growth phase, change the fresh medium, add MTX, the effect concentration is 11uM, and routinely culture in the incubator with 5% CO2 and 37°C, rinse with PBS 3 times after 24 hours, add no Continue culturing in the medium of MTX until the surviving cells in the culture flask grow to about 80% for passage. At this time, the sensitive cells die and float up, while the insensitive cells continue to survive, but the surviving cells grow slowly and change their shape;

[0019] (3)...

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Abstract

The invention relates to two human choriocarcinoma multidrug resistant cell lines. The preservation numbers of the human choriocarcinoma multidrug resistant cell lines are CCTCC NO: C2018245 or CCTCCNO: C2018246. Human choriocarcinoma JAR and JEG3 cell lines are selected as parental cells, the action concentration of a chemotherapy drug methotrexate (MTX) is gradually increased from 11 micro moleper liter to 88 micro mole per liter to establish JAR/MTX (CCTCC NO: C2018245) and JEG3/MTX (CCTCC NO: C2018246) drug-resistant cell lines. The JAR/MTX and JEG3/MTX can stably grow, passage and recover in the MTX of 88 micro mole per liter, and have the drug resistance indexes of 791.50 and 1040.04 to the MTX, and not only have drug resistance to the MTX, but also have cross drug resistance to VP-16, 5-FU, Taxol and ACTD. After the drug resistant cell lines are established, drug withdrawal culture is performed for 3 months, liquid nitrogen freeze saving is performed for half a year, the cellsrecover, and the drug resistance of the cells is redetected, the drug resistance of the two drug resistant cell lines separately accounts for 90% or above of the original drug resistance, and the stability of the drug resistance of the drug resistant cell lines is very good.

Description

technical field [0001] The invention relates to a drug-resistant cell line of human choriocarcinoma induced by methotrexate. Background technique [0002] Choriocarcinoma (choriocarcinoma for short) is a malignant tumor derived from embryonic trophoblast cells, which can be secondary to any type of pregnancy, including mole, miscarriage, and ectopic pregnancy or full-term pregnancy. The incidence of choriocarcinoma varies significantly around the world, and the incidence in Asia (approximately 1 case per 120-200 pregnancies) and Latin America is significantly higher than that in Europe and North America (approximately 1 case per 2000 pregnancies). The tumor has a high degree of malignancy, early and extensive metastasis, and its mortality rate can be as high as 90% before the advent of chemotherapy drugs. Because the tumor occurs in women of childbearing age, and most patients desire to preserve their reproductive function, chemotherapy rather than surgery is the main treat...

Claims

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Application Information

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IPC IPC(8): C12N5/09
Inventor 程晓东陈晓静吕卫国谢幸
Owner ATTACHED OBSTETRICS & GYNECOLOGY OSPITAL MEDICALCOLLEGE ZHEJIANG UNIV
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