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Application of STAT1 in treatment of paclitaxel-resistant epithelial ovarian cancer

A technology for paclitaxel and ovarian cancer, applied in the field of biomedicine, can solve the problems of STAT1 that have not been reported, and achieve the effect of novel guiding concept, reducing pressure, and good application prospects

Active Publication Date: 2020-05-22
JINSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is no report about the use of STAT1 of the present invention in the treatment of paclitaxel-resistant epithelial ovarian cancer

Method used

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  • Application of STAT1 in treatment of paclitaxel-resistant epithelial ovarian cancer
  • Application of STAT1 in treatment of paclitaxel-resistant epithelial ovarian cancer
  • Application of STAT1 in treatment of paclitaxel-resistant epithelial ovarian cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Construction of monoclonal paclitaxel-resistant epithelial ovarian cancer cell line

[0024] Build method:

[0025] Prepare human epithelial ovarian cancer cell line OVCAR-3 (USA, American Type Culture Collection Bank), paclitaxel (China, Sichuan Taiji Pharmaceutical Co., Ltd.). OVCAR-3 cell line is sensitive to paclitaxel, paclitaxel-resistant epithelial ovarian cancer cell line was constructed from OVCAR-3 cell line, the method is as follows:

[0026] (1) OVCAR-3 cells were inoculated into T25 culture flasks at a density of 30%, and cultured in RPMI-1640 (Sigma, USA) complete medium containing 10% fetal bovine serum (Gibco, USA). After 24 hours, the medium was discarded, washed with phosphate buffer, replaced with a complete medium containing 0.01 μM paclitaxel, and cultured for 24 hours. Then the culture medium was discarded, rinsed with phosphate buffered saline, replaced with paclitaxel-free complete medium for 3-5 days, and then replaced with 0.01 μM p...

Embodiment 2

[0029] Example 2 Construction of STAT1 function loss model

[0030] 1 Experimental method

[0031] 1.1 Cell culture

[0032] The cells used without special emphasis were cultured in culture plates or flasks using RPMI-1640 medium supplemented with 10% fetal bovine serum.

[0033] 1.2 Transfection of siRNA

[0034] Synthesized siRNA (synthesized by Shanghai Gemma Company) was shipped in the form of dry powder. Dissolve the dry powder with a certain volume of sterile DEPC water (supplied) to make a 20μM solution. The siRNA sequence is shown in Table 1:

[0035] Table 1

[0036]

[0037] Prepare well-grown cells to be treated, and transfect when the cells reach a suitable density. Firstly, 2 ml of Opti-MEM medium (USA, Gibco Company) was replaced in each well, and then an appropriate volume of transfection complex was prepared according to the needs of the experiment, and the transfection complex was added dropwise to each well in turn. For each well of a six-well plate...

Embodiment 3

[0043] Example 3 Construction of STAT1 function acquisition model

[0044] 1 Experimental method

[0045] 1.1 Cell culture

[0046]The cells used without special emphasis were cultured in culture plates or flasks using RPMI-1640 medium supplemented with 10% fetal bovine serum.

[0047] 1.2 Plasmid transfection

[0048] The structure of the STAT1 overexpression plasmid vector used is as follows: image 3 As shown, it was synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd., and DH5α Escherichia coli containing the plasmid vector was prepared.

[0049] When performing transfection experiments, first recover the DH5α Escherichia coli containing the STAT1 overexpression plasmid, and use the plasmid endotoxin-free plasmid mini-prep kit (China, Beijing Tiangen Company) for plasmid extraction. After obtaining the plasmid, configure it to 500ng / μl for use.

[0050] Prepare well-grown cells to be treated, and transfect when the cells reach a suitable density. Firstly, 2 ml of ...

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Abstract

The present invention relates to the technical field of biomedicines, specifically relates to an application of STAT1 in the treatment of paclitaxel-resistant epithelial ovarian cancer. According to the present invention, a monoclonal paclitaxel-resistant epithelial ovarian cancer cell line is constructed, and a STAT1 function deficient model is constructed by transfecting siRNA. The invention discloses that tumor stem cell-related markers in cells have increased expression level after expression of STAT1 reduces, indicating negative regulatory correlation between STAT1 and stemness of tumor stem cells. The STAT1 function model constructed by transfecting STAT1 overexpression plasmid shows reduced expression level of the tumor stem cell-related markers in the cells and reduced spheres formation of the cells after expression of STAT1 increases, indicating that overexpression of STAT1 can inhibit the stemness of drug-resistant cells. Moreover, the colony forming ability and proliferationgrowth ability of the cells also decreases, indicating that overexpression of STAT1 can inhibit the growth and proliferation of paclitaxel-resistant epithelial ovarian cancer cells.

Description

technical field [0001] The invention relates to the technical field of biomedicine, specifically, the use of STAT1 in the treatment of paclitaxel-resistant epithelial ovarian cancer. Background technique [0002] The incidence of ovarian cancer ranks third among female reproductive system malignancies, and it is also the most lethal gynecological tumor. The most common pathological type of ovarian cancer is epithelial ovarian cancer, and the current standard first-line treatment is tumor-reductive surgery combined with platinum-paclitaxel chemotherapy. [0003] Patients can obtain more obvious effects in the initial stage of treatment, and the remission rate of the initial treatment is high, but most patients relapse after two years of complete remission, and the relapse rate is as high as 80%. Recurrent epithelial ovarian cancer is resistant to almost all chemotherapy drugs, which greatly limits the clinical efficacy and application range of chemotherapy, and is also the m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P35/00
CPCA61K45/00A61P35/00
Inventor 许国雄王繁晨
Owner JINSHAN HOSPITAL FUDAN UNIV
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