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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, problems such as chemotherapy resistance still make its mortality rate the second highest among cancer deaths in women

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

Embodiment 1

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

[0174] The effect of miR-3074-5p transfection on the proliferation of ERα-positive breast cancer cells MCF-7 and T47D and ERα-negative breast epithelial cells MCF-10A was detected by automatic cell counter.

[0175] The three kinds of cells were collected and cultured by trypsinization respectively, counted by a cell counter and inoculated 3×10 4 , 4×10 4 , 3×10 4 cells / well, 0.5ml / well culture medium overnight. Use liposome transfection reagent lipofectamine2000 to transfect the three kinds of cells with miR-3074-5p and the negative control sequence respectively, after continuing to culture for 72 hours, digest and collect the cells in each well, and use trypan blue reagent with a final concentration of 0.1% Stain, read and compare the number of living cells in the negative control group and the miR-3074-5p overexpression group on an automatic cell counter, and use Graphpad Prism 6 for statisti...

Embodiment 2

[0177] Example 2 Effect of miR-3074-5p on the apoptosis rate of ERα-positive breast cancer cells

[0178]Annexin V-FITC / PI double staining kit was used to detect the effect of miR-3074-5p transfection on the apoptosis rate of ERα-positive breast cancer cell MCF-7.

[0179] Inoculate 5 × 10 in a 6-well plate 5 cells / well, 2ml culture medium / well, cultured overnight. Cells were transfected with miR-3074-5p mimic (miR-3074-5p mimic is the double-stranded form of endogenous miR-3074-5p mimic) and negative control sequence using lipofectamine2000, and cultured for 48 hours. After 1 hour, adjust slightly according to the instructions of the apoptosis detection kit for staining, wash the cells twice with PBS, add EDTA-free trypsin, digest and collect the cells in each well, centrifuge at 2000rpm for 5min, and resuspend with 1ml PBS. Counting, each group uniformly takes about 1×10 5 Cells were resuspended with 250ul of binding buffer in the kit, 2.5ul each of AnnexinV-FITC and PI w...

Embodiment 3

[0181] Example 3 The effect of tamoxifen on the expression level 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 cell MCF-7 was detected by fluorescent quantitative PCR technology.

[0183] MCF-7 cells were cultured for 48 hours using hormone-free conditions (10% activated charcoal-treated serum, phenol red-free medium), stimulated with 2 μM tamoxifen for 24 hours and 48 hours, washed once with ice-cold PBS, and collected in Trizol RNA was extracted, and reverse transcription was performed using specific primers for miR-3074-5p, and the expression level of miR-3074-5p was detected by fluorescent quantitative PCR.

[0184] The result is as image 3 As shown, the expression level of miR-3074-5p was significantly decreased after treatment of MCF-7 cells with 2 μM tamoxifen for 48 h, while there was no significant change after treatment for 24 h...

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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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