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Application of gnrh type Ⅱ antagonists in inhibiting the proliferation of progesterone-resistant endometrial cancer cells

A technology for endometrial cancer and endometrium, which is applied in the direction of antineoplastic drugs, drug combinations, and pharmaceutical formulations, and can solve problems such as downregulation of PR expression and progesterone resistance in endometrial cancer

Active Publication Date: 2019-03-26
PEOPLES HOSPITAL PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, long-term progesterone therapy can cause down-regulation of PR expression and lead to progesterone resistance in endometrial cancer

Method used

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  • Application of gnrh type Ⅱ antagonists in inhibiting the proliferation of progesterone-resistant endometrial cancer cells
  • Application of gnrh type Ⅱ antagonists in inhibiting the proliferation of progesterone-resistant endometrial cancer cells
  • Application of gnrh type Ⅱ antagonists in inhibiting the proliferation of progesterone-resistant endometrial cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Application of GnRH antagonists in inhibiting Ishikawa and Ishikawa-MPA cell proliferation

[0050] Cell lines in this example: Ishikawa-MPA cells and Ishikawa cells.

[0051] GnRH antagonists in this example: Cetrorelix and Trptorelix-1.

[0052] 1. Plank

[0053] (1) When the Ishikawa (ISK) and progesterone-resistant cell line (ISK-MPA) cells grow to the logarithmic phase, inoculate a 96-well plate;

[0054] (2) Use 0.25% trypsin solution (0.25g trypsin and 0.02g EDTA dissolved in 100ml PBS solution) to digest each cell in step (1) respectively, and then use DMEM-F12 high glucose containing 10% FBS The medium was digested and blown to make a single cell suspension, and 10 μl of the single cell suspension was counted with a cell counting plate, diluted with DMEM-F12 high glucose medium containing 10% FBS, so that the final concentration of each cell was 5000 cells / ml, add 100μl to each well;

[0055] (3) 5 duplicate wells for each type of cells, and 7 96...

Embodiment 2

[0068] Example 2. Application of GnRH antagonists in inhibiting xenograft tumors in nude mice

[0069] 1. Experimental materials

[0070] Ishikawa (ISK), progesterone-resistant cell line (Ishikawa-MPA), recipient animals 5-6 weeks old BALB / C genetic background female nude mice.

[0071] 2. Establishment of xenograft tumor model in nude mice

[0072] 1. Take endometrial cancer cells (Ishikawa, Ishikawa-MPA) in the logarithmic growth phase and prepare them into single cell suspensions (5×10 6 cells were suspended in 0.5ml of normal saline), to obtain Ishikawa single-cell suspension and Ishikawa-MPA single-cell suspension respectively;

[0073] 2. Under sterile conditions, the Ishikawa single-cell suspension and the Ishikawa-MPA single-cell suspension prepared in step 1 were respectively inoculated subcutaneously on the shoulders and backs of the bilateral forelimbs of five 5-6-week-old BALB / C genetic background female nude mice ( Each cell was inoculated with 5 mice);

[007...

Embodiment 3

[0090] Embodiment 3, the impact of GnRH antagonists on PR mRNA and GnRHR mRNA expression levels in different cell lines

[0091] Cell lines used in this example: Ishikawa (ISK), progesterone-resistant cell lines (Ishikawa-MPA, ISK-MPA). GnRH antagonists used in this example: Cetrorelix and Trptorelix-1.

[0092] 1. Plank

[0093] (1) When the Ishikawa (ISK) and progesterone-resistant cell line (ISK-MPA) cells grow to the logarithmic phase, inoculate a 96-well plate;

[0094] (2) Use 0.25% trypsin solution (0.25g trypsin and 0.02g EDTA dissolved in 100ml PBS solution) to digest each cell in step (1) respectively, and then use DMEM-F12 high glucose containing 10% FBS The medium was digested and blown to make a single cell suspension, and 10 μl of the single cell suspension was counted with a cell counting plate, diluted with DMEM-F12 high glucose medium containing 10% FBS, so that the final concentration of each cell was 5000 cells / ml, add 100μl to each well;

[0095] (3) 5 ...

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PUM

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Abstract

The invention discloses an application of a GnRH II type antagonist in inhibiting proliferation of progesterone-resistance endometrial cancer cells. Tests prove that the GnRH II type antagonist, when used independently or used in a mode of being combined with MPA, can be used for inhibiting the growth of the endometrial cancer cells, in particular the growth of the progesterone-resistance endometrial cancer cells; and by up-regulating PR expression, inhibiting a PI3K signal path and other mechanisms and by reversing progesterone resistance, the sensitivity of a progesterone-resistance endometrial cancer cell line to progesterone is enhanced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of GnRH type II antagonists in inhibiting the proliferation of progesterone drug-resistant endometrial cancer cells. Background technique [0002] Endometrial cancer is one of the three major malignant tumors of the female reproductive tract, accounting for 7% of female systemic malignant tumors and 25-30% of reproductive tract malignant tumors. In the past 10 to 20 years, the incidence of endometrial cancer is about twice that of the early 1970s, and the incidence tends to be younger. About 75% are early stage, and the lesion is still limited to the uterus at the time of diagnosis. [0003] In the treatment of endometrial cancer, in addition to conventional surgery, radiotherapy and chemotherapy, endocrine therapy has also become an important adjuvant therapy. At present, endocrine therapy is mostly used in the treatment of advanced and recurrent endomet...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K38/08A61P35/00A61K31/57
CPCA61K31/57A61K38/08A61K45/00A61K2300/00
Inventor 魏丽惠赵丽君李小平李明珠王建六
Owner PEOPLES HOSPITAL PEKING UNIV
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