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Application of long non-coding ribonucleic acid (lncRNA) in diagnosis and/or treatment of breast cancer

A technique for breast cancer, uses, applied in the field of molecular biology

Active Publication Date: 2018-07-13
WEIFANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the research on lncRNA is just in its infancy, and there is still a big gap in this field that researchers need to fill.

Method used

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  • Application of long non-coding ribonucleic acid (lncRNA) in diagnosis and/or treatment of breast cancer
  • Application of long non-coding ribonucleic acid (lncRNA) in diagnosis and/or treatment of breast cancer
  • Application of long non-coding ribonucleic acid (lncRNA) in diagnosis and/or treatment of breast cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Use Invitrogen's TRIzol to extract the total RNA in the cells.

[0035] 1.1 Aspirate the culture medium of normal breast epithelial cells MCF-10A or breast cancer cells MDA-MB-231, T47D, MCF-7 cells in the 6-well plate, add 1ml TRIzol to each well to make it cover the cells, then use a pipette or add The sample was pipetted 3 times, the cells should be completely lysed, and then transferred to a centrifuge tube.

[0036] 1.2 Add 0.2 mL of chloroform (0.2 mL of chloroform to 1 mL of TRIzol) into the centrifuge tube containing the lysate, shake and mix well on the shaker for 20 seconds, and place at room temperature for 5 minutes. Centrifuge at 12,000g at 4°C for 10 minutes, then pipette the upper aqueous phase containing total RNA into a new centrifuge tube. About 0.6mL of the upper aqueous phase can be absorbed per milliliter of TRIzol. The organic phase and interlayer contain DNA and proteins and should be avoided.

[0037] 1.3 Add isopropanol equal in volume to t...

Embodiment 2

[0050] Breast cancer cell MDA-MB-231 was infected with shRNA lentivirus, and Linc00426 in the cells was successfully knocked out, and a stable cell line was screened out. MDA-MB-231 cells infected with RNAi lentivirus were called SiLinc00426 / MDA-MB-231 , MDA-MB-231 cells infected with a control lentivirus called Scr / MDA-MB-231.

[0051] The Linc00426 shRNA lentivirus was customized by Shanghai Jikai Gene Company, and the carrier name is GV248. The sequence is: 5'-AAGGATGGAAATACAGAACAA-3', as shown in SEQ ID NO.4.

Embodiment 3

[0053] Wound healing assay to detect the migration ability of cells before and after infection: spread SiLinc00426 / MDA-MB-231 cells and Scr / MDA-MB-231 cells in a 35mm culture dish, after two days of growth, remove the conventional culture medium and replace it with 1 % fetal bovine serum culture medium. Use a 10 μL pipette tip to draw a uniform scratch on the bottom of the petri dish, and randomly select three fixed positions under an optical microscope to record the width of the scratch. The change of cell migration ability was judged according to the change of scratch width. The experiment was repeated three times, and the area of ​​the cell scratch was counted by ImageJ.

[0054] Experimental results such as image 3 As shown, knocking out Linc00426 can significantly inhibit the migration ability of breast cancer MDA-MB-231 cells.

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Abstract

The invention belongs to the field of molecular biology, and in particular relates to a long non-coding ribonucleic acid (lncRNA) breast cancer marker and knockout of lncRNA in application of treatment of breast cancer. The lncRNA is Linc00426, and the nucleotide sequence thereof is shown in SEQ ID NO. 1; the marker has a very significant up-regulation in breast cancer cells and can be used for screening the breast cancer in an auxiliary way. In addition, the knockout of the lncRNA can effectively inhibit the invasion, metastasis and proliferation of the breast cancer cells, thus being used for treating the breast cancer.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to the application of a lncRNA in the diagnosis and / or treatment of breast cancer. Background technique [0002] Breast cancer is one of the most common malignant tumors in women. Statistics at the end of the 20th century show that about 1.3 million people are diagnosed with breast cancer every year in the world, and 400,000 people die from the disease. Also in the last 10 years of the 20th century, the incidence of breast cancer in major cities such as Beijing, Tianjin, and Shanghai in mainland China has also ranked first and second among female malignant tumors. [0003] Although the current imaging diagnosis and pathological detection are highly accurate, they are often discovered at an advanced stage, which is extremely unfavorable for treatment. Therefore, how to realize the early diagnosis and effective treatment of breast cancer is an urgent need for breast cancer patients. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 尹崇高李洪利赵司顺李霞孙瑞美李佩瑞
Owner WEIFANG MEDICAL UNIV
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