具核梭杆菌通过靶向lncRNA NEAT1促进结肠癌的发展

陈渊文 张捷 沈火剑 朱宏毅 沈志勇

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微生物与感染 ›› 2022, Vol. 17 ›› Issue (5) : 282-289.

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具核梭杆菌通过靶向lncRNA NEAT1促进结肠癌的发展

陈渊文,张捷,沈火剑,朱宏毅,沈志勇

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Fusobacterium nucleatum promotes the development of colorectal carcinoma by targeting lncRNA NEAT1

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摘要

本研究旨在探讨具核梭杆菌对人结肠癌细胞增殖和凋亡的影响及其机制。取对数生长期人结肠癌HCT116细胞，设对照组、具核梭杆菌组（使用具核梭杆菌感染）、si-NEAT1组（转染si-NEAT1）组和具核梭杆菌+si-NEAT1组（转染si-NEAT1后使用具核梭杆菌感染）。利用qRT-PCR测量细胞中RNA的表达量。使用CCK-8实验检测细胞的增殖能力，流式细胞术计算细胞的凋亡率。凋亡相关蛋白caspase-3，Bcl-2和Bax的表达量通过western blot实验测定。具核梭杆菌感染结肠癌细胞后，结肠癌细胞的增殖能力上升，凋亡率降低，caspase-3和Bax蛋白表达量降低，而Bcl-2蛋白的表达量升高。此外，感染具核梭杆菌后结肠癌细胞中的NEAT1表达量上升。转染si-NEAT1后相比对照组，结肠癌细胞的增殖能力降低，凋亡率升高，caspase-3和Bax蛋白表达量提高，而Bcl-2蛋白的表达量下降。敲低NEAT1的表达对于具核梭杆菌影响结肠癌细胞增殖能力和凋亡率起到逆转作用。以上结果表明具核梭杆菌通过靶向上调NEAT1的表达促进结肠癌细胞的增殖并抑制结肠癌细胞的凋亡。

Abstract

The aim of this study was to investigate the effects of Fusobacterium nucleatum (F. nucleatum) on proliferation and apoptosis of human colorectal cancer (CRC) cells as well as its mechanism. Human CRC cells HCT116 at logarithmic growth stage were divided into control group, F. nucleatum group (infected with F. nucleatum), si-NEAT1 group (transfected with si-NEAT1) and F. nucleatum+si-NEAT1 group (infected with F. nucleatum after transfection with si-NEAT1). qRT-PCR was used to measure the level of RNA in cells. CCK-8 assay was performed to detect the proliferation of cells, while flow cytometry was used to calculate the apoptosis rate of cells. The expression levels of apoptosis related proteins caspase-3, Bcl-2 and Bax were determined by western blot. The proliferation of cells was enhanced, and the apoptosis rate of cells was downregulated. The expressions of caspase-3 and Bax were decreased, while the level of Bcl-2 was lifted. In addition, the expression of NEAT1 in CRC cells increased after infection with F. nucleatum. After transfection with si-NEAT1, the proliferation ability of CRC cells was decreased, the apoptosis rate was increased, and the expressions of caspase-3 and Bax protein were added, while the expression of Bcl-2 was reduced. Knockdown NEAT1 expression reversed the effect of F. nucleatum on proliferation and apoptosis of CRC cells. These results suggested that F. nucleatum could promote the proliferation and inhibit the apoptosis of CRC cells by up-regulating of NEAT1.

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陈渊文张捷沈火剑朱宏毅沈志勇. 具核梭杆菌通过靶向lncRNA NEAT1促进结肠癌的发展[J]. 微生物与感染. 2022, 17(5): 282-289

Fusobacterium nucleatum promotes the development of colorectal carcinoma by targeting lncRNA NEAT1[J]. Journal of Microbes and Infections. 2022, 17(5): 282-289

中图分类号： R735.3

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