Effects of T-cadherin silencing on ox-LDL induced biological behavior of human chorionic trophoblast cells HTR-8/SVneo

doi:10.11958/20220832

Abstract

Abstract:

Objective To investigate the effect of silencing T-cadherin on oxidized low density lipoprotein (ox-LDL) induced biological behavior of human chorionic trophoblast HTR-8/SVneo cells. Methods Human chorionic trophoblast cells HTR-8/SVneo were divided into the blank control group, the ox-LDL group, the T-cadherin small interfering RNA (siRNA) negative control group and the T-cadherin siRNA group. After each group was treated accordingly, the expression levels of T-cadherin messenger RNA (mRNA) in HTR-8/SVneo cells were detected by real-time fluorescence quantitative PCR in the four groups. The proliferation of cells in each group was detected by tetramethylazolate method. The clone formation of each group was detected by plate clone formation experiment. The apoptosis of each group was detected by flow cytometry. Transwell chamber test and scratch test were used to detect the invasion and migration ability of cells. The protein expression levels of Caspase-3, Bcl-2 related X protein (Bax), B cell lymphoma-2 (Bcl-2) and matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) of cells in each group were detected by Western blot assay. Results Compared with the blank control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were increased significantly in the ox-LDL group and the T-cadherin siRNA negative control group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were decreased significantly (P<0.05). Compared with the ox-LDL group and the T-cadherin siRNA negative control group, expression levels of T-cadherin mRNA, proliferation inhibition rate, apoptosis rate, Caspase-3 and Bax protein were decreased significantly in the T-cadherin siRNA group, and expression levels of clone formation rate, number of invasive cells, scratch healing rate, Bcl-2, MMP-2 and MMP-9 protein were increased significantly (P<0.05). Conclusion Silencing T-cadherin can inhibit ox-LDL induced abnormal apoptosis of HTR-8/SVneo cells, and promote cell proliferation, invasion and migration.

Key words: cadherins, lipoproteins, IDL, trophoblasts, cell proliferation, apoptosis, T-cadherin, oxidized low density lipoprotein

CLC Number:

R714.43

Figures/Tables 8

References 23

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组别	细胞增殖抑制率	细胞克隆形成率
空白对照组	10.96±1.68	39.26±4.35
ox-LDL组	32.64±4.11^a	15.42±2.37^a
T-cadherin siRNA阴性对照组	31.23±3.97^a	16.85±2.16^a
T-cadherin siRNA组	20.60±3.06^bc	28.08±3.18^bc
F	55.031^＊＊	75.487^＊＊

组别	细胞增殖抑制率	细胞克隆形成率
空白对照组	10.96±1.68	39.26±4.35
ox-LDL组	32.64±4.11^a	15.42±2.37^a
T-cadherin siRNA阴性对照组	31.23±3.97^a	16.85±2.16^a
T-cadherin siRNA组	20.60±3.06^bc	28.08±3.18^bc
F	55.031^＊＊	75.487^＊＊

组别	侵袭细胞数（个/视野）	划痕愈合率（%）
空白对照组	140.22±22.95	25.66±4.30
ox-LDL组	65.47±8.04^a	9.08±1.09^a
T-cadherin siRNA阴性对照组	66.06±8.06^a	9.12±1.13^a
T-cadherin siRNA组	98.53±10.03^bc	20.11±3.77^bc
F	39.563^＊＊	46.731^＊＊

组别	侵袭细胞数（个/视野）	划痕愈合率（%）
空白对照组	140.22±22.95	25.66±4.30
ox-LDL组	65.47±8.04^a	9.08±1.09^a
T-cadherin siRNA阴性对照组	66.06±8.06^a	9.12±1.13^a
T-cadherin siRNA组	98.53±10.03^bc	20.11±3.77^bc
F	39.563^＊＊	46.731^＊＊

组别	Caspase-3	Bax	Bcl-2	MMP-2	MMP-9
空白对照组	0.96±0.09	1.01±0.10	1.03±0.09	1.02±0.11	0.97±0.12
ox-LDL组	1.65±0.18^a	1.70±0.18^a	0.21±0.02^a	0.26±0.04^a	0.31±0.04^a
T-cadherin siRNA阴性对照组	1.62±0.17^a	1.72±0.19^a	0.18±0.03^a	0.28±0.04^a	0.29±0.03^a
T-cadherin siRNA组	1.17±0.14^bc	1.35±0.13^bc	0.66±0.07^bc	0.73±0.08^bc	0.72±0.07^bc
F	31.227^＊＊	28.419^＊＊	274.909^＊＊	150.516^＊＊	120.541^＊＊