Establishment and preliminary analysis of lung adenocarcinoma cell lines with stable MINK1 knockdown

Zhang Tao, Zhang Lian, Zhang Chundong

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Journal of Chongqing Medical University ›› 2024, Vol. 49 ›› Issue (09) : 1072-1078. DOI: 10.13406/j.cnki.cyxb.003589
Basic research

Establishment and preliminary analysis of lung adenocarcinoma cell lines with stable MINK1 knockdown

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Abstract

Objective To establish lung adenocarcinoma cell lines with stable knockdown of the expression of encoding Misshapen/NIK-related kinase（MINK1）and analyze the effects of stable MINK1 knockdown on cell proliferation，migration，and cisplatin treatment. Methods In lung adenocarcinoma A549 and PC-9 cells，cell lines with stable MINK1 knockdown were constructed based on the pLKO.1-shRNA lentiviral expression system. The knockdown efficiency was detected at the mRNA and protein levels using qRT-PCR and Western blot. CCK-8 and Transwell assays were used to assess the effects of stable MINK1 knockdown on cell proliferation and migration. Flow cytometry was used to detect changes in cell cycle and apoptosis. The sensitivity of cells with stable MINK1 knockdown to cisplatin treatment was analyzed. The level of DNA damage in cells was detected by immunofluorescence staining of the DNA damage marker γH2AX. Results qRT-PCR and Western blot showed that shRNA significantly downregulated the expression of MINK1. Proliferation and migration were reduced in lung adenocarcinoma cells with stable MINK1 knockdown. MINK1 knockdown significantly enhanced the cytotoxic effect of the chemotherapy drug cisplatin on cancer cells and induced an increase in the level of DNA damage. Conclusion Lung adenocarcinoma cell lines with stable MINK1 knockdown were successfully established using A549 and PC-9 cells. Stable MINK1 knockdown can inhibit cancer cell proliferation and migration and may reduce the drug resistance of lung cancer cells by regulating the DNA damage repair process.

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lung adenocarcinoma / MINK1 / cell proliferation / cell migration / DNA damage repair

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Zhang Tao , Zhang Lian , Zhang Chundong. Establishment and preliminary analysis of lung adenocarcinoma cell lines with stable MINK1 knockdown. Journal of Chongqing Medical University. 2024, 49(09): 1072-1078 https://doi.org/10.13406/j.cnki.cyxb.003589

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