Human ovarian cancer multidrug resistant cell line

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

Active Publication Date: 2013-07-31
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 ovarian cancer multi-

Method used

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  • Human ovarian cancer multidrug resistant cell line
  • Human ovarian cancer multidrug resistant cell line
  • Human ovarian cancer multidrug resistant cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Induction and establishment of human ovarian cancer multidrug-resistant cell line HO-8910 / VP16

[0021] The human ovarian cancer multidrug-resistant cell line HO-8910 / VP16 was established by gradually increasing the concentration of VP16 and intermittently induced in vitro. The specific steps are as follows:

[0022] (1) Culture medium for human ovarian cancer cell line HO-8910 (RPMI-1640 medium containing 10% calf serum), 5% CO2, 37°C, culture to 70%-90% adherence rate, Remove the supernatant, add VP161000ng / ml impact culture for 1 hour, remove VP16100ng / 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 expanded to 70%-90% adherence rate, repeat the shock culture with VP16100ng / ml culture solution for 1 hour and 9 times, that is, complete 10 shock cultures with fluorouracil and 1000ng / ml paclitaxel culture solution for 1 hour, an...

Embodiment 2

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

[0026] The multidrug-resistant human ovarian cancer cell line HO-8910 / VP16 in the logarithmic growth phase was taken, and the cell morphology was observed under an inverted microscope. The results were 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 the cell growth curves of HO-8910 and HO-8910 / VP16, see figure 1 .

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Abstract

According to the invention, human ovarian cancer cell line HO-8910 is adopted as an induction object; with a high-dose impact method, and with an in-vitro induction method with gradually increased VP16 concentration and intermittent action, the human ovarian cancer multidrug resistant cell line HO-8910/VP16 is established. The cell ling is preserved at China General Microbiological Culture Collection Center on December 24th, 2012, and has a preservation number of CGMCC NO.7052. The cell line provided by the invention has typical multidrug resistance. With the cell line, an experiment basis is provided for further researching resistance reversal approaches.

Description

technical field [0001] The invention belongs to the technical field of cell engineering and relates to a human cell line, in particular to a human ovarian cancer multidrug-resistant cell line and a method for establishing the same. Background technique [0002] Among malignant tumors of female reproductive organs, ovarian cancer ranks third in incidence and first in mortality. This is because most ovarian cancers are insidious and lack effective and accurate early diagnosis methods. About 70% to 80% of patients are diagnosed at an advanced stage. [0003] Chemotherapy is one of the important means to treat human malignant tumors. However, the drug resistance of tumor cells often limits the further improvement of the curative effect, and finally makes the treatment fail. Tumor multidrug resistance (multidrug resistance, MDR) is an important cellular defense mechanism for tumor cells to avoid drug attack. The phenomenon of cross-resistance. Overcoming the multidrug resistan...

Claims

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Application Information

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IPC IPC(8): C12N5/09
Inventor 不公告发明人
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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