Let-7a过表达对结肠黏液腺癌细胞LS-174T增殖及侵袭性的影响

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Abstract Objective: To study the effect of let-7a overexpression on the proliferation and invasiveness of colonic mucinous adenocarcinoma cell line LS-174T. Methods: The let-7a overexpression lentiviral vector was constructed and infected with colonic mucinous adenocarcinoma LS-174T cells. A blank control group was established: no virus transfection; B transfected empty virus group; C1 transfection group (concentration 1× 106 TU/L): transfection of hsa-let-7a [containing let-7a virus and enhanced green fluorescent protein (EGFP)]; C2 transfection group (concentration 1×107 TU/L); C3 transfection group (concentration 1×108 TU/L). The infection efficiency of each group was observed by fluorescence microscope. The expression of let-7a was detected by RT-qPCR. The proliferation of each groupcells was detected by MTT assay. The invasion and migration ability of cells were evaluated by Transwell invasion and scratch test. RT-qPCR and Western blot analysis of TGF-β1 mRNA and protein levels were expressed. Results: The expression of green fluorescence protein in each group was observed by inverted fluorescence microscope. The fluorescence expression of C3 transfection group was the strongest, with transfection efficiency over 90%, which was positively correlated with the initial virus concentration (P<0.05). The expression levels of let-7a in each group were detected by RT-qPCR. The results showed that the expression level of let-7a in the transfected group was significantly higher than that in the negative control group, and the expression level of let-7a in the transfected cells was positively correlated with the gradient of lentivirus concentration (P<0.05). The results of Western blot and RT-qPCR showed that the expression of TGF-β1 mRNA and protein decreased gradually after let-7a overexpression (P<0.05). The results of invasion and scratching showed that the overexpression of let-7a reduced the invasion and migration ability of LS-174T cells, and was negatively correlated with the initial virus concentration (P<0.05). Conclusion: Overexpression of let-7a can inhibit the invasion and metastasis of LS-174T cells by down-regulating the content of TGF-β1.

Key words： let-7a invasion and metastasis epithelial-mesenchymal transition colonic mucinous aden-
ocarcinoma

Received: 20 February 2019

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	HUANG Chongjie
	CAO Weilan
	HONG Huang
	LIU Pengpeng
	LIU Changbao

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2019/V49/I10/724

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