Human ovarian cancer drug-resistant cell strain and culture method thereof

A technology for drug-resistant cells and ovarian cancer, applied in the direction of tumor/cancer cells, tissue culture, animal cells, etc., can solve the problems of drug resistance and characteristics that are very different

Inactive Publication Date: 2008-09-03
李红霞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, resistance to these drugs is often encountered
At present, the research on drug resistance mostly uses the concentration gradient method to ind

Method used

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  • Human ovarian cancer drug-resistant cell strain and culture method thereof
  • Human ovarian cancer drug-resistant cell strain and culture method thereof
  • Human ovarian cancer drug-resistant cell strain and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] SKOV3 cells were induced with topotecan (TOP) drug, and SKOV3 / TOP drug-resistant cell line was established.

[0031] Drugs: Topotecan (TOP) is a product of Guizhou Hanfang Pharmaceutical Co., Ltd.

[0032] (1) Dosage: according to the formula [(60×D) / (5000)]×2×10 3 , where 60 is the average adult body weight (kg), D clinical dosage (mg.kg -1 .d -1 ), 5000 is the average adult blood volume (ml), 2 is divided by the hematocrit 50%, 10 3 Replace mg with ug, refer to the clinical chemotherapy dose for ovarian cancer patients, TOP 1.2mg / m 2 , taking the dosage of clinical high-dose pulse therapy as 3 times the usual dose as the standard, the final concentration of TOP is 2160ng / ml as the high-dose pulse dosage;

[0033] (2) Parental cell culture: human ovarian epithelial cancer cell line SKOV3 cells were grown in RPMI1640 medium containing 15% calf serum, placed in 5% CO 2 , Cultured in a 37°C incubator.

[0034] (3) Induction of ovarian cancer drug-resistant cell line...

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Abstract

The invention relates to a TOP medicament resistant cell line human ovarian cancer, with a accession number of CGMCC No 1217, which is selecting human ovarian cancer cell line as parental cell, putting the cell into RPMI1640 culture medium containing 15% calf serum, putting into incubator with 5% CO2 and 37 DEG C to culture; acquiring cells in logarithmic growth phase, counting after digesting with 0.25% pancreatin, inoculating 5*105/ml into culture bottle according to amount of 5ml in each bottle, when cells adheres to wall after 24 hours, reacting for 2 hours with a TOP final content of 2160ng/ml, discarding culture medium, washing by phosphate buffer for 3 times, replacing fresh culture medium, second impulsing after growing is resumed, and repeatedly operating, until it is impulsed with medicament for 13 times. The cell line is constructed for 10 months, medicament resistance of cell is 10.67 per cell, P<0.01 comparing with sensitive cell, has cross resistance to mitoxantrone, camptothecin, vincristine besides resistance to TOP.

Description

technical field [0001] The present invention relates to a method of selecting human ovarian cancer cell line SKOV3 as the parent cell, selecting topotecan, an anticancer drug commonly used in clinical practice, as an inducer, and intermittently administering it in the form of repeated high-dose shocks to establish the topotecan-resistant cell line SKOV3 / TOP Drug-resistant human ovarian cancer cell line and its culture method. Background technique [0002] Ovarian cancer is one of the most lethal gynecological malignancies. At present, the combination of paclitaxel and platinum is the first-line chemotherapy for ovarian cancer, and the combination of topotecan and platinum is the second-line chemotherapy. However, resistance to these drugs is often encountered. At present, the research on drug resistance mostly uses the concentration gradient method to induce drug-resistant cell lines in vitro, but this method is far from the characteristics of repeated and intermittent drug...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/09
Inventor 李红霞周友珍张素梅魏丽惠
Owner 李红霞
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