miR-126-3p promotes the proliferation, migration and invasion of ovarian cancer cells by targeting PIK3R1

doi:10.3877/cma.j.issn.2095-1221.2024.01.003

Abstract

Abstract:

Objective

To investigate the role of miR-126-3p/PIK3R1 axis proliferation, migration and invasion cells in ovarian cancer.

Methods

miR-126-3p mimic, miR-126-3p inhibitor and PIK3R1 plasmid were transfected into SKOV3 and A2780 ovarian cancer cells. Cell proliferation/migration was assessed by CCK-8, scratch and Transwell assays. Western blot and qPCR were used to detect the expression levels of PIK3R1 and miR-126-3p. A Dual-luciferase reporter assay was used to detect the target gene of miR-126-3p, PIK3R1. An independent samle t-test was used to compare between two groups, a one-way analysis of variance was used to compare among multiple groups and an LSD-t test was used for pairwise comparison among various groups.

Results

Compared with the normal ovarian epithelial cell line IOSE80, the expression levels of miR-126-3p were up-regulated in ovarian cancer cell lines SKOV3, A2780, HO-8910 [ (2.30 ± 0.18) , (1.86 ± 0.11) , (1.26 ± 0.12) vs (1.00 ± 0.07) ] (all P < 0.05) ; Inhibition of miR-126-3p expression inhibited the proliferation [SKOV3 cells : 72 h (0.94 ± 0.13) vs (1.14 ± 0.08) , A2780 cells: 72 h (0.83 ± 0.09) vs (1.11 ± 0.12) ], migration [ (23.00 ± 6.08) % vs (37.67 ± 6.43) %, (25.67 ± 4.04) % vs (40.00 ± 6.58) %], invasion of SKOV3, A2780 cells[ (97.00 ± 17.35) vs (134.33 ± 13.32) , (97.00 ± 7.00) vs (127.00 ± 9.17) cells] (all P < 0.05) . And dual luciferase report experiment verified miR-126-3p targeted PIK3R1 gene in SKOV3 and A2780 cells. After overexpression of miR-126-3p, the expression of PIK3R1 protein was down-regulated in SKOV3 and A2780 cells [ (0.50 ± 0.08) vs (1.01 ± 0.04) , (0.54 ± 0.03) vs (1.00 ± 0.03) ] (all P < 0.001) . Compared with miR-126-3p mimic, miR-126-3p mimic+pPIK3R1 inhibited the proliferation [SKOV3 cells: 72 h (1.04 ± 0.14) vs (1.55 ± 0.12) , A2780 cells: 72 h (0.87 ± 0.09) vs (1.32 ± 0.11) ], migration ability[ (27.00 ± 2.00) %vs (34.00 ± 2.00) %, (24.67 ± 3.22) %vs (33.00 ± 4.00) %], invasion ability of SKOV3 and A2780 cells [ (60.67 ± 7.64) vs (135.00 ± 6.00) , (63.33 ± 10.02) vs (125.67 ± 9.87) cells], the protein expression of PIK3R1 was increased [ (1.92 ± 0.16) vs (1.00 ± 0.11) , (3.15 ± 0.06) vs (0.99 ± 0.12) ] (all P < 0.05) .

Conclusion

miR-126-3p promotes the proliferation, migration and invasion of ovarian cancer cells by targeting PIK3R1.

Key words: Ovarian cancer, MiR-126-3p, PIK3R1, Proliferation, Migration

wei Wei, Yina Wang, Xiang Kong. miR-126-3p promotes the proliferation, migration and invasion of ovarian cancer cells by targeting PIK3R1[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(01): 19-26.

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URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2024.01.003

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Figures/Tables 14

References 21

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基因	序列（5'→3'）	预期扩增长度（bp）
PIK3R1	上游TGGTGGACGGCGAAGTAAAG	150
PIK3R1	下游CATTGAGGGAGTCGTTGTGCTG	150
β-actin	上游TGCTGTCCCTGTATGCCTCTG	223
β-actin	下游TTGATGTCACGCACGATTTCC	223
miR-126-3p	上游CGCGCGCGCGACCGGA	107
miR-126-3p	下游AGTGCAGGGTCCGAGGTATT	107
U6	上游CTCGCTTCGGCAGCACA	102
U6	下游AACGCTTCACGAATTTGCGT	102

基因	序列（5'→3'）	预期扩增长度（bp）
PIK3R1	上游TGGTGGACGGCGAAGTAAAG	150
PIK3R1	下游CATTGAGGGAGTCGTTGTGCTG	150
β-actin	上游TGCTGTCCCTGTATGCCTCTG	223
β-actin	下游TTGATGTCACGCACGATTTCC	223
miR-126-3p	上游CGCGCGCGCGACCGGA	107
miR-126-3p	下游AGTGCAGGGTCCGAGGTATT	107
U6	上游CTCGCTTCGGCAGCACA	102
U6	下游AACGCTTCACGAATTTGCGT	102

分组	实验次数	迁移率（%）		侵袭数目（个）
分组	实验次数	SKOV3细胞	A2780细胞	SKOV3细胞	A2780细胞
对照	3	37.67 ± 6.43	40.00 ± 6.58	134.33 ± 13.32	127.00 ± 9.17
miR-126-3p inhibitor	3	23.00 ± 6.08	25.67 ± 4.04	97.00 ± 17.35	97.00 ± 7.00
t值		2.870	3.223	2.957	4.506
P值		0.045	0.032	0.041	0.011

分组	实验次数	迁移率（%）		侵袭数目（个）
分组	实验次数	SKOV3细胞	A2780细胞	SKOV3细胞	A2780细胞
对照	3	37.67 ± 6.43	40.00 ± 6.58	134.33 ± 13.32	127.00 ± 9.17
miR-126-3p inhibitor	3	23.00 ± 6.08	25.67 ± 4.04	97.00 ± 17.35	97.00 ± 7.00
t值		2.870	3.223	2.957	4.506
P值		0.045	0.032	0.041	0.011

分组	实验次数	SKOV3-PIK3R1蛋白	A2780-PIK3R1蛋白
对照	3	1.01 ± 0.04	1.00 ± 0.03
miR-126-3p mimic	3	0.50 ± 0.08	0.54 ± 0.03
t值		10.252	17.997
P值		< 0.001	< 0.001

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