Preparation method of miR-126 transfected ovarian cancer cell line

A technology of 1mir-126 and 2mir-126, which is applied in the field of culturing ovarian cancer cell lines, can solve the problems of unclear expression and role of ovarian cancer

Inactive Publication Date: 2014-03-05
ZHEJIANG UNIV
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Problems solved by technology

MicroRNA plays a very important role in the process of tumorigenesis. miR-126 is a member of the microRNA family. There are a large number of literature reports that miR-126 is closely related to malignant tumors such as lung cancer, gastric cancer, breast cancer, and colon cancer. The expression and role in ovarian cancer are not yet clear

Method used

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Embodiment 1

[0036] Embodiment 1, a method for preparing an ovarian cancer cell line transfected with miR-126, the following steps are performed in sequence:

[0037] 1) Parental cell culture:

[0038] Human ovarian mucinous cystadenocarcinoma cell line SKOV3, at 37 ° C, 5% (volume %) CO 2 Cultured in DMEM high-glucose medium containing 10% FBS and 1% Anti-Anti;

[0039] Then, digest and subculture with PBS containing 0.25% trypsin and 0.02% EDTA, once every 2-3 days;

[0040] The above-mentioned DMEM high-sugar culture solution containing 10% FBS and 1% Anti-Anti is: add 100 ml of FBS (fetal calf serum, fetal bovine serum) and 10 ml of Anti-Anti ( Antibiotic-Antimycotic, antibacterial-antifungal agent).

[0041] The above PBS containing 0.25% trypsin and 0.02% EDTA is as follows: add 0.25g trypsin + 0.02g EDTA (ethylenediaminetetraacetic acid) to every 100ml of PBS buffer (pH7.4).

[0042] Remarks: The above passage only needs to be done once, and the early passage cells with good gro...

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Abstract

The invention discloses a preparation method of an miR-126 transfected ovarian cancer cell line. The preparation method comprises the following steps: 1, culture of parental cells: conducting subculturing on a human ovarian mucinous cystadenocarcinoma cell line SKOV3; 2, transfection preparation of miR-126: using a lentiviral technology, inoculating the SKOV3 cells in logarithmic phase obtained in the step 1 into a 6 cm petri dish, adding 2 ml of a whole culture medium, and washing medium cells again with 2 ml of a serum-free medium when cells grow to 80%-90% fusion 24 hours later, and getting ready for transfection; and 3, arrangement of an miR-126 transfection overexpression group. The miR-126 transfection overexpression group has the following purposes: observation on biological behavior changes of ovarian cancer cells after miR-126 transfection; and research on effect of miR-126 in occurrence and development of ovarian cancer and searching for new targets for clinical treatment of ovarian cancer.

Description

technical field [0001] The invention relates to a culture method for selecting a human ovarian cancer cell line SKOV3 as a parental cell and using a lentiviral vector to prepare an ovarian cancer cell line transfected with miR-126. Background technique [0002] Ovarian cancer is the gynecological malignancy with the highest mortality rate. Most of them are in the advanced stage and have been widely metastasized to the abdominal cavity and pelvic organs. Surgery, chemotherapy, and radiotherapy can only prolong the survival time of patients to the maximum extent. In terms of the current level of medical development It is difficult to cure completely. Due to the high degree of malignancy and recurrence rate of ovarian cancer, coupled with the tendency to develop drug resistance, its 5-year survival rate is only about 20%. Therefore, it is an urgent clinical need to study the mechanism of occurrence and development of ovarian cancer and find the target for its treatment. Micro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
Inventor 周建维王双燕费菁赵家耀李建琼王红亚
Owner ZHEJIANG UNIV
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