Micro RNA (Ribonucleic Acid) targeting estrogen receptor alpha and anti-tumor application of micro RNA

A tiny, useful technology, applied in the field of medicine, can solve the problem of high mortality due to chemotherapy resistance, and achieve the effect of improving the inhibitory effect

Pending Publication Date: 2021-07-20
上海市生物医药技术研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the clinical prognosis and five-year survival rate of breast cancer have improved significantly, proble

Method used

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  • Micro RNA (Ribonucleic Acid) targeting estrogen receptor alpha and anti-tumor application of micro RNA
  • Micro RNA (Ribonucleic Acid) targeting estrogen receptor alpha and anti-tumor application of micro RNA
  • Micro RNA (Ribonucleic Acid) targeting estrogen receptor alpha and anti-tumor application of micro RNA

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0173] Example 1 Effect of MiR-3074-5P on ERα positive and ERα negative cells

[0174] Transfection of MIR-3074-5P to ERα-positive breast cancer cells MCF-7, T47D and ERα negative breast epithelial cell MCF-10A proliferation effect were detected by ERα positive.

[0175] Three cells were collected by trypsin digestion, and the counts were counted and 3 × 10, respectively. 4 , 4 × 10 4 , 3 × 10 4 Cell / well, 0.5 ml / well medium culture overnight. The MIR-3074-5P and negative control sequences were transfected with the MiR-3074-5P and negative control sequences with liposomal transfection reagent, and the cells were continued for 72 hours, and the cells in each well were collected, and the final concentration was 0.1%. Dyeing and read and compare the negative control group and the number of live cells expressed in the MIR-3074-5P group were subjected to a statistical and analyzed by GraphPad Prism 6.

[0176] Such as figure 1 As shown, overexpression of MIR-3074-5P significantly i...

Example Embodiment

[0177] Effect of Example 2 MiR-3074-5P on the apoptosis rate of breast cancer cells in ERα positive

[0178]The effect of transfection of MIR-3074-5P on ERα positive breast cancer cells MCF-7 apoptosis rate was detected by Annexin V-FITC / PI Double Dye Kit.

[0179] Inoculated in 6-well plates 5 × 10 5 Cell / well, 2 ml medium / well, overnight culture. Cells were transfected with MiR-3074-5P MIMIC (MiR-3074-5P MIMIC) and the negative control sequences using liposome-3074-5p mimic (MIR-3074-5P MIMIC) and the negative control sequences, respectively, and continued to cultivate 48 After the hour, the stain was taken for staining with the apoptosis test kit specification, and the cells were washed twice with PBS, and the cells containing non-EDTA were added, and the cells in each well were collected, 2000 rpm, 5 min centrifuge, and resuspended with 1 ml of PBS. Count, each group is unified to take about 1 × 10 5 The cells were resuspended in a 250 ul kit, and annexinv-FITC, PI was a...

Example Embodiment

[0181] Example 3 Effect of Tamoxifen on Expression of MiR-3074-5P in ERα positive breast cancer cells

[0182] The effect of a certain amount of tamoxifen stimulation on the expression level of MIR-3074-5P in ERα positive breast cancer cells was detected by fluorescence quantitative PCR technology.

[0183] After 48 hours of cultured MCF-7 cells (serum-free medium treatment of 10% activated carbon treatment), 2 μm tamoxifen stimulated 24h, 48h, wash the cells once with ice PBS, collected in Trizol And extract RNA, the anti-transcription is performed using a specific primer of MiR-3074-5P, and the expression level of MIR-3074-5P was detected by fluorescence quantitative PCR technology.

[0184] Such as image 3 As shown, after 2 μm tamoxifen, MCF-7 cells were treated 48 h, and the expression level of miR-3074-5P was significantly reduced, and there was no significant change in the treatment 24h ( image 3 ). This result suggests that TAM effects induces a significant reduction in exp...

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Abstract

The invention relates to an application of micro RNA (Ribonucleic Acid) in tumor resistance. More specifically, the invention relates to an application of micro RNA-3074-5p in resisting estrogen receptor alpha (ER alpha) positive tumors, and an application of micro RNA-3074-5p in chemotherapy sensitization. The micro RNA-3074-5p disclosed by the invention can inhibit proliferation of ER[alpha] positive breast cancer cells and expression levels of ER[alpha] and anti-apoptotic protein Bcl-2 in the breast cancer cells, induce apoptosis of the ER[alpha] positive breast cancer cells, and enhance the anti-tumor effect of chemotherapeutic drugs, thereby providing a new way and means for effectively treating ER[alpha] positive tumors.

Description

technical field [0001] The invention belongs to the field of medicine, and more specifically relates to a microRNA targeting estrogen receptor alpha and its antitumor application. Background technique [0002] Breast cancer is the most common malignant tumor in women worldwide, and the incidence of breast cancer in Chinese women was about 15% in 2015. Although the clinical prognosis and five-year survival rate of breast cancer have been significantly improved, problems such as chemotherapy resistance still make its mortality rate the second highest in female cancer mortality. About 70% of breast cancers in situ express estrogen receptor α (ERα). ERα is not only the best prognostic indicator for breast cancer treatment, but also a key target molecule for endocrine therapy of breast cancer. ERα, as a nuclear transcription factor, can mediate the proliferation of tumor cells stimulated by estrogen. ERα is highly expressed in breast cancer, endometrial cancer and ovarian cance...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61K31/712A61K31/7125A61K31/138A61K45/06A61P35/00A61K49/00
CPCA61K31/7088A61K31/712A61K31/7125A61K31/138A61K45/06A61P35/00A61K49/0008A61K2300/00
Inventor 顾文文张璇杨龙甄兴兴顾一骅王健
Owner 上海市生物医药技术研究院
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