抑制PTBP1对脉络膜黑色素瘤细胞增殖和侵袭力的影响

doi:10.3969/j.issn.1673-8640.2023.01.010

摘要/Abstract

摘要：

目的探讨多聚嘧啶区结合蛋白1（PTBP1）对脉络膜黑色素瘤（CM）细胞增殖活性和迁移、侵袭能力的影响。方法选取人CM细胞系OCM-1，根据转染序列的不同分为si-PTBP1组（转染PTBP1特异性干扰序列）、si-Control组（转染阴性对照序列）和空白组（不作任何处理）。分别检测各组细胞PTBP1 mRNA的表达，以及PTBP1、上皮型钙黏蛋白（E-Cad）和波形蛋白的表达。通过细胞实验分析各组细胞的增殖活性和迁移、侵袭能力。结果 si-PTBP1组PTBP1 mRNA相对表达量显著低于si-Control组和空白组（P<0.05），si-Control组和空白组之间差异无统计学意义（P>0.05）。培养24、48、72和96 h，si-PTBP1组细胞增殖活性均低于si-Control组和空白组（P<0.05）。si-PTBP1组迁移细胞数和侵袭细胞数均少于si-Control组和空白组（P<0.05）。si-PTBP1组PTBP1和波形蛋白相对表达量低于si-Control组和空白组（P<0.05），E-Cad相对表达量高于si-Control组和空白组（P<0.05）。结论抑制OCM-1细胞PTBP1表达可抑制细胞增殖活性，削弱细胞的迁移和侵袭能力，其机制可能与抑制上皮-间质转化有关。

关键词: 多聚嘧啶区结合蛋白1, 细胞增殖, 细胞侵袭, 上皮-间质转化, 脉络膜黑色素瘤

Abstract:

Objective To investigate the effect of polypyrimidine tract-binding protein 1（PTBP1） on the proliferation activity，migration and invasion abilities of choroidal melanoma（CM） cells. Methods Human CM cell line OCM-1 was selected and classified into si-PTBP1 group（transfected with PTBP1 specific interference sequence），si-Control group（transfected with negative control sequence） and blank group（without any treatment），according to different transfection sequences. The expressions of PTBP1 mRNA，PTBP1，E-cadherin（E-Cad） and vimentin protein were determined. The proliferation activity，migration and invasion abilities were determined by cytological experiment. Results The relative expression level of PTBP1 mRNA in si-PTBP1 group was lower than those in si-Control group and blank group（P<0.05），and there was no statistical significance between si-Control group and blank group（P>0.05）. At 24，48，72 and 96 h of culture，the proliferation activities in si-PTBP1 group were lower than those in si-Control group and blank group（P<0.05）. The number of migrating cells and the number of invasive cells in si-PTBP1 group were lower than those in si-Control group and blank group（P<0.05）. The relative expression levels of PTBP1 and vimentin protein in si-PTBP1 group were lower than those in si-Control group and blank group（P<0.05），and the relative expression level of E-Cad was higher than those in si-Control group and blank group（P<0.05）. Conclusions The inhibition of the expression of PTBP1 in OCM-1 can inhibit cell proliferation activity and reduce migration and invasion，and its mechanism may be related to the inhibition of epithelial-mesenchymal transformation.

Key words: Polypyrimidine tract-binding protein 1, Cell proliferation, Cell invasion, Epithelial-mesenchymal transformation, Choroidal melanoma

中图分类号:

R446.1

尚贞君, 魏威, 刘瑞菡. 抑制PTBP1对脉络膜黑色素瘤细胞增殖和侵袭力的影响[J]. 检验医学, 2023, 38(1): 51-55.

SHANG Zhenjun, WEI Wei, LIU Ruihan. Effect of inhibiting PTBP1 on the proliferation and invasiveness of choroidal melanoma cells[J]. Laboratory Medicine, 2023, 38(1): 51-55.

图/表 7

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组别	培养12 h	培养24 h	培养48 h	培养72 h	培养96 h
si-PTBP1组	0.62±0.11	0.75±0.12^*#	1.13±0.09^*#	1.26±0.11^*#	1.42±0.10^*#
si-Control组	0.63±0.04	1.01±0.10	1.29±0.11	1.50±0.08	1.79±0.05
空白组	0.64±0.10	0.96±0.05	1.34±0.14	1.47±0.12	1.76±0.12
F值	0.116	12.608	5.783	9.505	29.893
P值	0.892	0.001	0.014	0.002	<0.001

组别	培养12 h	培养24 h	培养48 h	培养72 h	培养96 h
si-PTBP1组	0.62±0.11	0.75±0.12^*#	1.13±0.09^*#	1.26±0.11^*#	1.42±0.10^*#
si-Control组	0.63±0.04	1.01±0.10	1.29±0.11	1.50±0.08	1.79±0.05
空白组	0.64±0.10	0.96±0.05	1.34±0.14	1.47±0.12	1.76±0.12
F值	0.116	12.608	5.783	9.505	29.893
P值	0.892	0.001	0.014	0.002	<0.001

组别	迁移细胞数	侵袭细胞数
si-PTBP1组	110.83±8.84^*#	91.50±3.94^*#
si-Control组	130.00±7.69	120.17±9.85
空白组	134.67±14.32	123.33±5.24
F值	8.386	39.558
P值	0.004	<0.001

组别	迁移细胞数	侵袭细胞数
si-PTBP1组	110.83±8.84^*#	91.50±3.94^*#
si-Control组	130.00±7.69	120.17±9.85
空白组	134.67±14.32	123.33±5.24
F值	8.386	39.558
P值	0.004	<0.001

组别	PTBP1	E-Cad	波形蛋白
si-PTBP1组	0.21±0.05^*#	0.75±0.09^*#	0.25±0.08^*#
si-Control组	0.83±0.06	0.33±0.04	0.69±0.03
空白组	0.80±0.07	0.36±0.09	0.66±0.06
F值	155.291	54.509	97.964
P值	<0.001	<0.001	<0.001