lncRNA FTX silencing inhibits gastric cancer cell proliferation through the miR-22-3p/NLRP3 inflammasome pathway

doi:10.3760/cma.j.cn371439-20220429-00040

Abstract

Abstract:

Objective To investigate the regulatory effect of long non-coding RNA （lncRNA） FTX on gastric cancer cell proliferation through miR-22-3p/NOD-like receptor protein 3 （NLRP3） inflammasome pathway. Methods The gastric cancer cell line NCI-N87 were divided into blank control group， si-FTX-NC group， si-FTX group， si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group. Quantitative real-time fluorescent PCR was performed to analyze the expression levels of lncRNA FTX and miR-22-3p， clone formation assay was performed to analyze the proliferation ability of NCI-N87 cells， western blotting was performed to analyze the expressions of NLRP3 inflammasome pathway proteins， and dual-luciferase reporter assay was performed to analyze the targeting relationship between lncRNA FTX and miR-22-3p. Results The relative expressions of lncRNA FTX in the blank control group， si-FTX-NC group， si-FTX group， si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group were 1.03±0.09， 1.01±0.15， 0.42±0.08， 0.45±0.06 and 0.46±0.13 respectively， with a statistically significant difference （F=52.19， P<0.001）. The relative expressions of miR-22-3p were 1.04±0.12， 0.97±0.08， 2.26±0.15， 2.23±0.13 and 1.15±0.11 respectively， with a statistically significant difference （F=178.53， P<0.001）. Compared with the blank control group and si-FTX-NC group， the relative expressions of lncRNA FTX in the si-FTX group， si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group decreased （all P<0.001）. Compared with the blank control group， si-FTX-NC group and si-FTX+miR-22-3p inhibitor group， the relative expressions of miR-22-3p in the si-FTX group and si-FTX+miR-22-3p inhibitor-NC group increased （all P<0.001）. The clones of the five groups were 115.50±7.25， 112.33±8.46， 54.83±5.17， 56.17±6.32 and 85.67±9.43， with a statistically significant difference （F=91.67， P<0.001）. The levels of NLRP3 protein in the five groups were 1.84±0.17， 1.86±0.12， 0.95±0.09， 0.97±0.11 and 1.28±0.19， with a statistically significant difference （F=60.62， P<0.001）. Compared with the blank control group and si-FTX-NC group， the number of clones and the level of NLRP3 protein of the si-FTX group， si-FTX+miR-22-3p inhibitor-NC group and si-FTX+miR-22-3p inhibitor group decreased （all P<0.05）. Compared with the si-FTX+miR-22-3p inhibitor group， the number of clones and the level of NLRP3 protein in the si-FTX group and si-FTX+miR-22-3p inhibitor-NC group decreased （all P<0.05）. The dual-luciferase reporter assay found that miR-22-3p was the target gene of lncRNA FTX. Conclusion Silencing the expression of lncRNA FTX can inhibit the proliferation of gastric cancer cells， and the mechanism may be related to the regulation of lncRNA FTX on the miR-22-3p/NLRP3 inflammasome pathway.

Key words: Stomach neoplasms, RNA, long noncoding, MicroRNAs, Cell proliferation

Quan Zhenhao, Xu Feipeng, Huang Zhe, Huang Xianjin, Chen Rihong, Sun Kaiyu, Hu Xu, Lin Lin. lncRNA FTX silencing inhibits gastric cancer cell proliferation through the miR-22-3p/NLRP3 inflammasome pathway[J]. Journal of International Oncology, 2023, 50(4): 202-207.

Figures/Tables 8

References 14

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细胞株	FTX	miR-22-3p
GES-1	1.02±0.04	0.99±0.05
NCI-N87	3.45±0.14^ab	0.37±0.09^a
SNU-1	2.98±0.15^a	0.46±0.10^a
HGC-27	3.26±0.11^ab	0.43±0.09^a
KATO Ⅲ	3.31±0.17^ab	0.39±0.08^a
F值	362.60	58.05
P值	<0.001	<0.001

组别	lncRNA FTX	miR-22-3p
空白对照组	1.03±0.09	1.04±0.12
si-FTX-NC组	1.01±0.15	0.97±0.08
si-FTX组	0.42±0.08^ab	2.26±0.15^abc
si-FTX+miR-22-3p inhibitor-NC组	0.45±0.06^ab	2.23±0.13^abc
si-FTX+miR-22-3p inhibitor组	0.46±0.13^ab	1.15±0.11
F值	52.19	178.53
P值	<0.001	<0.001

组别	活力（%）	克隆数量（个）
空白对照组	100.00±9.72	115.50±7.25
si-FTX-NC组	100.00±8.51	112.33±8.46
si-FTX组	55.62±7.13^abc	54.83±5.17^abc
si-FTX+miR-22-3p inhibitor-NC组	52.68±7.34^abc	56.17±6.32^abc
si-FTX+miR-22-3p inhibitor组	75.39±8.46^ab	85.67±9.43^ab
F值	46.09	91.67
P值	<0.001	<0.001

组别	NLRP3	caspase-1/pro-caspase-1	消皮素D-N/消皮素D	IL-1β/pro-IL-1β
空白对照组	1.84±0.17	0.73±0.05	0.49±0.04	0.65±0.09
si-FTX-NC组	1.86±0.12	0.72±0.08	0.51±0.06	0.63±0.08
si-FTX组	0.95±0.09^abc	0.41±0.04^abc	0.26±0.03^abc	0.34±0.05^abc
si-FTX+miR-22-3p inhibitor-NC组	0.97±0.11^abc	0.43±0.06^abc	0.24±0.05^abc	0.31±0.04^abc
si-FTX+miR-22-3p inhibitor组	1.28±0.19^ab	0.67±0.05	0.39±0.06^ab	0.55±0.07
F值	60.62	45.58	54.78	33.04
P值	<0.001	<0.001	<0.001	<0.001

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