Effect of lncRNA TINCR on the biological behavior of trophoblast HTR-8/SVneo cells and its mechanism

doi:10.3877/cma.j.issn.2095-1221.2024.03.006

Abstract

Abstract:

Objective

To investigate the effect of lncRNA tissue differentiation-inducing differentiation inducing non-protein coding RNA (TINCR) on proliferation, migration, invasion, epithelial-stromal transformation and apoptosis of trophoblast HTR-8/SVneo cells.

Methods

HTR-8/SVneo cells were cultured in vitro and then divided into three groups, which included the control group (standard culture) , negative group (transfected NC-siRNA) and interference group (transfected TINCR-siRNA) respectively. The expression level of TINCR in the cells was detected by real-time fluorescence quantitative PCR, cell proliferation was detected by CCK-8 method, the migration of HTR-8/SVneo cells was detected by scratch healing assay, and cell migration and cell invasion were tested by Transwell chamber. The expression of epithelial-mesenchymal transformation (EMT) related proteins E-cadherin, N-cadherin, Vimentin and Wnt/β-catenin key proteins Wnt5a and β-catenin in cells were checked by Western blotting.

Results

Compared with the control group and the negative group, the expression level of TINCR (0.25 ± 0.03 vs 1.00 ± 0.08; 0.25 ± 0.03 vs 0.98 ± 0.06) was decreased in the interference group; compared with the control group and negative group, the expression levels of Vimentin (0.92 ± 0.08 vs 0.33 ± 0.05, 0.92 ± 0.08 vs 0.31 ± 0.04) and N-cadherin (0.87 ± 0.07 vs 0.29 ± 0.04) . 0.87 ± 0.07 vs 0.32 ± 0.05) were increased in HTR-8/SVneo cells in interference group, while the expression level of E-cadherin protein were decreased; compared with the control group and negative group, the expression level of Wnt5a (0.97 ± 0.06 vs 0.48 ± 0.04, 0.97 ± 0.06 vs 0.46 ± 0.03) and β-catenin proteins (0.89 ± 0.05 vs 0.37 ± 0.03, 0.89 ± 0.05 vs 0.39 ± 0.03) were increased in interference group (P < 0.05) .

Conclusion

Down-regulating TINCR expression may promote the proliferation, migration, invasion and epithelial-mesenchymal transformation of HTR-8/SVneo cells and inhibit their apoptosis by activating the Wnt/β-catenin signaling pathway.

Key words: Preeclampsia, lncRNA TINCR, Cell proliferation, Migration, Wnt/β- catenin signaling pathway

Bo Li, Xiuyan Ma, Jie Sun. Effect of lncRNA TINCR on the biological behavior of trophoblast HTR-8/SVneo cells and its mechanism[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2024, 14(03): 167-172.

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Figures/Tables 9

References 27

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基因	引物序列（5'→3'）	预期扩增产物长度（bp）
TINCR	上游TGTGGCCCAAACTCAGGGATACAT	216
	下游AGATGACAGTGGCTGGAGTTGTCA
GAPDH	上游CATGTTCGTCATGGGTGTGAACCA	132
	下游AGTGATGGCATGGACTGTGGTCAT

基因	引物序列（5'→3'）	预期扩增产物长度（bp）
TINCR	上游TGTGGCCCAAACTCAGGGATACAT	216
	下游AGATGACAGTGGCTGGAGTTGTCA
GAPDH	上游CATGTTCGTCATGGGTGTGAACCA	132
	下游AGTGATGGCATGGACTGTGGTCAT

分组	实验次数	24 h	48 h	72 h
对照	3	0.36 ± 0.03	0.54 ± 0.04	0.83 ± 0.05
阴性对照	3	0.35 ± 0.03	0.52 ± 0.03	0.85 ± 0.06
TINCR-siRNA	3	0.37 ± 0.03	0.76 ± 0.06^ab	1.23 ± 0.09^ab
F值		1.000	78.492	96.592
P值		0.383	< 0.001	< 0.001

分组	实验次数	24 h	48 h	72 h
对照	3	0.36 ± 0.03	0.54 ± 0.04	0.83 ± 0.05
阴性对照	3	0.35 ± 0.03	0.52 ± 0.03	0.85 ± 0.06
TINCR-siRNA	3	0.37 ± 0.03	0.76 ± 0.06^ab	1.23 ± 0.09^ab
F值		1.000	78.492	96.592
P值		0.383	< 0.001	< 0.001

分组	实验次数	划痕愈合率（%）	迁移细胞数（个）	侵袭细胞数（个）
对照	3	46.35 ± 5.02	85.56 ± 6.75	67.38 ± 5.23
阴性对照	3	44.81 ± 4.76	90.25 ± 7.42	62.52 ± 5.17
TINCR-siRNA	3	65.40 ± 5.91^ab	138.85 ± 11.35^ab	105.46 ± 8.85^ab
F值		42.900	102.453	112.753
P值		< 0.001	< 0.001	< 0.001

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