Application of miRNA-1293 in preparation of anti-colorectal tumor drug

The technology of mirna-1293 and 1. mirna-1293 is applied in the application field of preparing anti-colorectal tumor drugs, and can solve the problems of insensitivity to chemotherapy drugs, harmful and unhelpful, tumor recurrence, etc., and achieve the effect of high-efficiency anti-tumor effect.

Active Publication Date: 2019-11-12
THE SIXTH AFFILIATED HOSPITAL OF 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, and more than half of the patients need to receive chemotherapy before and after surgery. However, many clinical retrospective analyzes have pointed out that a large number of patients who receive chemotherapy are actually insensitive to chemotherapy drugs or even harmful. , and even developed tumor drug resistance
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  • Application of miRNA-1293 in preparation of anti-colorectal tumor drug
  • Application of miRNA-1293 in preparation of anti-colorectal tumor drug
  • Application of miRNA-1293 in preparation of anti-colorectal tumor drug

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Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1 colorectal tumor cell culture

[0040] Human colorectal tumor cells HCT 116 and RKO (purchased from ATCC, the American Type Culture Collection) were cultured in RPMI-1640 and DMEM medium containing 10% FBS at 37°C, 5% CO 2 , saturated humidity in a carbon dioxide incubator. When the cell confluence reached 80%-90%, the cells were digested and passaged with 0.25% trypsin.

Embodiment 2

[0041] Example 2 Cell Transfection

[0042] 1. use RNAiMAX transfection reagent transfected HCT 116 and RKO cells with miRNA-1293mimics and negative control NC mimics respectively:

[0043] The specific sequences of miRNA-1293mimics and negative control NC mimics (purchased from Gemma Gene Company) are as follows:

[0044] NC mimics:

[0045] SEQ ID NO.1 5'-UUCUCCGAACGUGUCACGUTT-3'

[0046] SEQ ID NO.2 5'-ACGUGACACGUUCGGAGAATT-3'

[0047] miR-1293mimics:

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

[0049] SEQ ID NO.4 5'-ACAAAUCUCCAGACCACCAUU-3'

[0050] 2. The specific experimental steps are as follows:

[0051] (1) Digest well-growing HCT 116 and RKO cells in the logarithmic growth phase with trypsin, and press HCT 116 cells per well at 3×10 5 3×10 RKO cells per well 5 The cell density of each was planted in a 6-well plate, cultured in RPMI-1640 and DMEM medium, and placed in a carbon dioxide incubator for 24 hours;

[0052] (2) After culturing overnight, di...

Embodiment 3

[0055] Example 3 Q-PCR detection of the expression level of miRNA-1293

[0056] After transfecting HCT 116 cells and RKO cells (cells obtained in step (4) in Example 2) with NC mimics and miRNA-1293mimics, RNA was extracted, reversed by tailing, and its expression level was detected by Q-PCR. Specific steps are as follows:

[0057] 1. Collect HCT 116 and RKO cells 48 hours after transfection with Trizol reagent, and extract RNA:

[0058] (1) 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 The lysate was transferred into a 1.5mL centrifuge tube and left at room temperature for 5 minutes;

[0059] (2) Add 200 μL of chloroform to each tube, mix well, and let stand at room temperature for 3 minutes;

[0060] (3) Centrifuge at 4°C and 12000rpm for 10min, and transfer the upper aqueous phase to another new 1.5mL cen...

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Abstract

The invention belongs to the field of biomedicine and particularly relates to application of miRNA-1293 or miRNA-1293 mimics or miRNA-1293 accelerants in preparation of an anti-colorectal tumor drug.The invention finds for the first time that the miRNA-1293 can inhibit proliferation of colorectal tumor cells and promote apoptosis of the colorectal tumor cells, which has important significance inreducing the death rate of colorectal tumor.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of miRNA-1293 or its mimic or its promoter 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. The latest data show that there are approximately 1.4 million new cases of colorectal tumors worldwide each year, and nearly 700,000 people die of colorectal tumors. The incidence rate of colorectal tumors in Guangzhou is 36.81 / 100,000, and an average of 7.5 people are diagnosed with colorectal tumors every day. And with the improvement of quality of life and the change of eating habits, more and more patients with colorectal tumors. [0003] In addition, compared with Western countries, the early diagnosis rate of colorectal tumors in ...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P35/00C12Q1/6886
CPCA61K31/7105A61P35/00C12Q1/6886C12Q2600/178C12Q2600/158
Inventor 黄春颖林梦梦赵绪平杨湘玲陈家和汪中扬刘焕亮
Owner THE SIXTH AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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