Effects of circ_0000218 on the proliferation, migration and invasion of cervical cancer HeLa cells by sponging miR-1182

doi:10.3877/cma.j.issn.2095-1221.2021.02.007

Abstract

Abstract:

Objective

To explore whether circular RNA 0000218 (circ_0000218) sponges miR-1182 to regulate the proliferation, migration and invasion of cervical cancer HeLa cells.

Methods

Expression levels of circ_0000218 and miR-1182 in the cancer tissues and paracancer tissues from 43 patients with cervical cancer were analyzed by real-time quantitative PCR (RT-qPCR) . Cervical cancer cells HeLa were divided into si-NC group, si-circ_0000218 group, miR-NC group, miR-1182 group, si-circ_0000218+anti-miR-NC group, and si-circ_0000218+anti-miR-1182 group. The cell counting kit (CCK-8) method and Transwell experiment were used to analyze the effects of circ_0000218 and miR-1182 expression on the proliferation, migration and invasion of HeLa cells. Western blotting was used for detecting the expression of Ki-67, matrix metalloproteinase-2 (MMP-2) and MMP9 proteins. The dual luciferase reporter assay and RT-qPCR were applied to validate the targeting relationship between circ_0000218 and miR-1182. Pared t test was used to compare the difference of the expression of circ_0000218 and miR-1182 between cancer tissues and paracancer tissues. Independenr sample t test was used to compare the difference between groups.

Results

The expression of circ_0000218 in the cervical cancer tissue was significantly higher than that in the adjacent tissues (4.17±0.32 vs 1.00±0.05, P < 0.001) , while the expression of miR-1182 was significantly lower than that in the adjacent tissues (0.33±0.03 vs 1.00±0.05, P < 0.001) . Compared with the si-NC group, the HeLa cell proliferation activity (0.86±0.04 vs 0.37±0.03) , migration (86.73±7.13 vs 38.52±3.19) and invasion (66.80±4.95 vs 26.58±2.55) , and the expression levels of Ki-67 (0.57±0.05 vs 0.18±0.02) , MMP-2 (0.74±0.07 vs 0.28±0.03) and MMP-9 (0.64±0.04 vs 0.22±0.02) proteins in the si-circ_0000218 group were significantly decreased (P < 0.001) . Compared with the miR-NC group , the proliferation (0.88±0.04 vs 0.46±0.04) , migration (89.74±5.53 vs 46.63±3.79) and invasion (68.03±4.34 vs 34.63±3.37) of HeLa cells, and the expression of Ki-67 (0.59±0.04 vs 0.24±0.02) , MMP-2 (0.76±0.05 vs 0.33±0.03) and MMP-9 (0.66±0.04 vs 0.29±0.03) proteins in the miR-1182 group were decreased significantly. Circ_0000218 targeted and negatively regulated miR-1182 expression (P < 0.001) . Compared with the si-circ_0000218+ anti-miR-NC group, the HeLa cell proliferation activity (0.35±0.03 vs 0.76±0.04) , migration (35.58±3.11 vs 77.04±4.08) and invasion (25.44±2.29 vs 57.61±3.47) , and the expression levels of Ki-67 (0.16±0.02 vs 0.46±0.04) , MMP-2 (0.26±0.02 vs 0.65±0.04) and MMP-9 (0.20±0.02 vs 0.57±0.04) proteins in the si-circ_0000218+ anti-miR-1182 group were significantly increased (P < 0.001) .

Conclusion

circ_0000218 can promote the proliferation, migration and invasion of cervical cancer HeLa cells by sponging miR-1182.

Key words: Circular RNA, Cervical cancer, Cell proliferation, Migration, Invasion, miR-1182

Jing Guo, Weihong Qi. Effects of circ_0000218 on the proliferation, migration and invasion of cervical cancer HeLa cells by sponging miR-1182[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2021, 11(02): 113-119.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2021.02.007

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Figures/Tables 14

References 18

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临床病理参数		例数
年龄（岁）
	<50	18
	≥50	25
临床分期
	Ⅰ	6
	Ⅱ	11
	Ⅲa	5
	Ⅲb	9
	Ⅳ	12
分化程度
	高	13
	中	12
	低	18
淋巴结转移
	有	16
	无	27

临床病理参数		例数
年龄（岁）
	<50	18
	≥50	25
临床分期
	Ⅰ	6
	Ⅱ	11
	Ⅲa	5
	Ⅲb	9
	Ⅳ	12
分化程度
	高	13
	中	12
	低	18
淋巴结转移
	有	16
	无	27

基因	序列（5'-3'）		预期扩增产物（bp）
circ_0000218	F: TGGAGTGCAGCTTGAAGGTTTAT	R: TTCAGAGCCAAAGCGGGAGA	258
circ_0000218	RT: CGTTTCTTCCAAAATCTGTATTTCGGGCTCGTTAACAACAACTCCTTA		258
GAPDH	F: GGGAAACTG TGGCGTGAT	R: GGGTGTCGCTGTTGAAGT	280
miR-1182	F: ACTTGTCACTGCCTGTCTCC	R: CCAACAGTCACATCCCTCCC	150
miR-1182	RT: CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCAGCTGC		150
U6	F: CTCGCT TCGGCAGCACA	R: AACGCTTCACGAATTTGCGT	108

基因	序列（5'-3'）		预期扩增产物（bp）
circ_0000218	F: TGGAGTGCAGCTTGAAGGTTTAT	R: TTCAGAGCCAAAGCGGGAGA	258
circ_0000218	RT: CGTTTCTTCCAAAATCTGTATTTCGGGCTCGTTAACAACAACTCCTTA		258
GAPDH	F: GGGAAACTG TGGCGTGAT	R: GGGTGTCGCTGTTGAAGT	280
miR-1182	F: ACTTGTCACTGCCTGTCTCC	R: CCAACAGTCACATCCCTCCC	150
miR-1182	RT: CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCAGCTGC		150
U6	F: CTCGCT TCGGCAGCACA	R: AACGCTTCACGAATTTGCGT	108

分组	实验次数	WT-circ_0000218	MUT-circ_0000218
miR-NC	3	1.04±0.06	1.02±0.08
miR-1182	3	0.56±0.04	1.01±0.06
t值		11.529	0.173
P值		< 0.001	0.871

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