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Application of miRNA-5571 in preparation of drugs for resisting colorectal tumor

A miRNA-5571, 1. The technology of miRNA-5571 is applied in the application field of miRNA-5571 in the preparation of anti-colorectal tumor drugs, which can solve the problems of limiting therapeutic effect and tumor recurrence

Active Publication Date: 2019-07-05
SUN YAT SEN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the treatment of colorectal tumors is mainly based on surgery, supplemented by radiotherapy and chemotherapy. Drug treatment includes chemotherapy and targeted drug therapy. However, once tumor cells develop drug resistance, chemotherapy drugs cannot fully exert their anti-tumor effects. Even if most of the tumor cells are killed, this small part of drug-resistant tumor cells will continue to grow and cause tumor recurrence. Therefore, problems such as tumor drug resistance greatly limit the use of colorectal cancer chemotherapy. treatment effect

Method used

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  • Application of miRNA-5571 in preparation of drugs for resisting colorectal tumor
  • Application of miRNA-5571 in preparation of drugs for resisting colorectal tumor
  • Application of miRNA-5571 in preparation of drugs for resisting colorectal tumor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1.Q-PCR detection of the expression level of miR-5571-5p

[0046] After HCT 116 cells and SW 480 cells were transfected with NC / miR-5571-5p mimics, RNA was extracted, reversed by tailing, and its expression was detected by Q-PCR. NC / miR-5571-5pmimics (double-stranded RNA fragments) in this experiment were purchased from Gemma Genetics. The sequences are

[0047] NC mimics:

[0048] SEQ ID NO.5 5'-UUCUCCGAACGUGUCACGUTT-3'

[0049] SEQ ID NO.6 5'-ACGUGACACGUUCGGAGAATT-3'

[0050] miR-5571-5p mimics:

[0051] SEQ ID NO.2 5'-CAAUUCUCAAAGGAGCCUCCC-3'

[0052] SEQ ID NO.7 5'-GAGGCUCCUUUGAGAAUUGUU-3'

[0053] Experimental steps:

[0054] 1. Collect HCT 116 and SW 480 cells 48 hours after transfection with Trizol reagent, and extract RNA.

[0055] 2. Suck off the medium in the well plate, wash 2-3 times with PBS, remove the residual medium as much as possible, then add 1ml Trizol reagent to each well to lyse the cells, pipette continuously until clear, and then ...

Embodiment 2

[0075] Example 2.miR-5571-5p inhibits the proliferation ability of colorectal tumor cells

[0076] After HCT 116 cells and SW 480 cells were transfected with NC / miR-5571-5p mimics, the cell proliferation ability was analyzed by xCELLigence RTCA DP real-time cell analysis system.

[0077] Experimental steps:

[0078] After adding 100 μl of medium to the wells of the E-Plate 16 plate, place the E-Plate 16 plate on the RTCA DP instrument, check the baseline, and ensure that the contact of all wells is normal (the Cell Index of all wells is lower than 0.063.).

[0079] 1. Take out the E-Plate 16 plate, digest and inoculate the transfected HCT 116 and SW 480 cells on the E-Plate 16 plate, the inoculum volume of HCT 116 and SW 480 cells is 3000 and 4000 respectively.

[0080] 2. Place the E-Plate 16 in the incubator for 30 minutes.

[0081] 3. Place the E-Plate 16 plate with added cells on the RTCA DP instrument, and observe continuously for 7 days in the incubator for real-time d...

Embodiment 3

[0085] Example 3. MiR-5571-5p inhibits the migration ability of colorectal tumor cells

[0086] HCT 116 cells and SW 480 cells were transfected with NC / miR-5571-5p mimics, respectively, and the migration ability of cells was verified by transwell chamber migration assay.

[0087] Experimental steps:

[0088] After digesting the transfected HCT 116 and SW 480 cells, resuspend the cells with serum-free medium, centrifuge to remove the medium, then resuspend the cells with serum-free medium, count the cells, and adjust the cell concentration to 5×10 6 / ml.

[0089] 1. Add 100 μl of HCT 116 and SW 480 cell suspensions to the chambers on the transwell plate, and place the transwell plate in the incubator for 30 minutes.

[0090] 2. Add 600 μl of medium containing 10% serum to the lower chamber of the transwell plate.

[0091] 3. Place the transwell plate in the incubator for 24 hours.

[0092] 4. After 24 hours, the cells were fixed with formaldehyde, and stained with Coomassie...

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Abstract

The invention belongs to the field of biological medicines, and relates to an application of miRNA-5571 or a mimic thereof or an accelerant thereof in the preparation of drugs for resisting colorectaltumor. The research finds that by transfecting miR-5571 or the mimics thereof, the migration and invasion capacity of colorectal tumor cells can be effectively inhibited. The invention finds the inhibition effect of the miR-5571 on the colorectal tumor for the first time, and the miR-5571 has important significance in reducing the death rate of the colorectal tumor.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of miRNA-5571 in the preparation of anti-colorectal tumor drugs. Background technique [0002] Colorectal tumor is one of the most common malignant tumors in the world, with extremely high morbidity and mortality, and has become an important public problem that seriously threatens public health. According to the 2016 China Cancer Statistics Report, colorectal tumors are the top five most common cancers, and the incidence rate is still on the rise in the past 10 years. Although the diagnosis and treatment technology of tumors has been continuously improved in recent years, the situation of diagnosis and treatment of colorectal tumors in my country is still very severe. The 5-year survival rate of colon cancer is only 57.6%, and that of rectal cancer is only 56.9%, which is lower than that of neighboring East Asian countries and European and American countries. ...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K45/06A61K48/00A61K31/713A61K31/7088A61P35/00
CPCA61K31/7088A61K31/713A61K45/00A61K45/06A61P35/00
Inventor 林梦梦黄春颖杨湘玲黄兰兰温创宇陈骏雄刘瑞贤汪中扬刘焕亮
Owner SUN YAT SEN UNIV
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