MUC16对骨肉瘤细胞增殖和侵袭能力的影响

doi:10.3969/j.issn.1673-8640.2022.010.014

摘要/Abstract

摘要：

目的探讨黏蛋白16（MUC16）对人骨肉瘤细胞系U2OS增殖和侵袭能力的影响。方法将U2OS细胞按转染序列的不同分为sh-MUC16组（转染MUC16干扰序列）、sh-Control组（转染阴性对照序列）和空白组（不处理）。采用实时荧光定量聚合酶链反应（PCR）检测各组MUC16 mRNA表达，噻唑蓝（MTT）法检测细胞增殖活性，Transwell法检测细胞迁移和侵袭能力，免疫印迹法检测细胞中MUC16、上皮型钙黏蛋白（E-Cad）和神经型钙黏蛋白（N-Cad）表达。结果 sh-MUC16组MUC16 mRNA相对表达量显著低于sh-Control组和空白组（P<0.05），sh-Control组与空白组之间差异无统计学意义（P>0.05）。sh-MUC16组培养24、48、72和96 h时细胞增殖活性低于sh-Control组和空白组（P<0.05）。sh-MUC16组迁移和侵袭细胞数均低于sh-Control组和空白组（P<0.05）。sh-MUC16组MUC16和N-Cad相对表达量均低于sh-Control组和空白组（P<0.05），E-Cad相对表达量高于sh-Control组和空白组（P<0.05）。结论沉默U2OS细胞中MUC16基因表达可抑制细胞增殖、迁移和侵袭，其机制可能与抑制上皮-间质转化有关。

关键词: 黏蛋白16, 骨肉瘤, 细胞增殖, 细胞侵袭, 上皮-间质转化

Abstract:

Objective To investigate the influence of mucin 16（MUC16） on the proliferation and invasion of human osteosarcoma cell line U2OS. Methods The U2OS cells were classified into sh-MUC16 group（transfected with MUC16 interference sequence），sh-Control group（transfected with negative control sequence） and blank group（without any treatment）. Real-time fluorescence quantitative polymerase chain reaction（PCR） was used to determine the expression of MUC16 mRNA in each group，methyl thiazolyl tetrazolium（MTT） method was used to determine cell proliferation activity，Transwell method was used to determine cell migration and invasion ability，and western blotting was used to determine the expressions of MUC16 epithelial E-cadherin（E-Cad） and neuropathic cadherin（N-Cad） in cells. Results The relative expression level of MUC16 mRNA in sh-MUC16 group were lower than those in sh-Control group and blank group（P<0.05），while there was no statistical significance between sh-Control group and blank group（P>0.05）. The cell proliferation activities in sh-MUC16 group at 24，48，72 and 96 h were lower than those in sh-Control group and blank group（P<0.05）. The number of migrating and invasive cells in sh-MUC16 group were lower than those in sh-Control group and blank group（P<0.05）. The relative expression levels of MUC16 and N-Cad in sh-MUC16 group were lower than those in sh-Control group and blank group，while the relative expression level of E-Cad was higher than those in sh-Control group and blank group（P<0.05）. Conclusions Silencing the expression of MUC16 gene in U2OS cells can inhibit cell proliferation activity，cell migration and invasion，and its mechanism may be related to the inhibition of epithelial-mesenchymal transition.

Key words: Mucin 16, Osteosarcoma, Cell proliferation, Cell invasion, Epithelial-mesenchymal transition

李名武, 王勤志, 孙法瑞. MUC16对骨肉瘤细胞增殖和侵袭能力的影响[J]. 检验医学, 2022, 37(10): 979-983.

LI Mingwu, WANG Qinzhi, SUN Farui. Influence of MUC16 on the cell proliferation and invasion of osteosarcoma cells[J]. Laboratory Medicine, 2022, 37(10): 979-983.

图/表 7

参考文献 19

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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
sh-MUC16组	0.15±0.06	0.20±0.07^*#	0.25±0.07^*#	0.36±0.08^*#	0.50±0.05^*#
sh-Control组	0.18±0.05	0.34±0.07	0.39±0.03	0.57±0.04	0.75±0.07
空白组	0.15±0.03	0.35±0.11	0.42±0.12	0.63±0.10	0.73±0.05
F值	0.873	6.170	5.087	19.958	35.958
P值	0.438	0.011	0.021	<0.001	<0.001

组别	培养12 h	培养24 h	培养48 h	培养72 h	培养96 h
sh-MUC16组	0.15±0.06	0.20±0.07^*#	0.25±0.07^*#	0.36±0.08^*#	0.50±0.05^*#
sh-Control组	0.18±0.05	0.34±0.07	0.39±0.03	0.57±0.04	0.75±0.07
空白组	0.15±0.03	0.35±0.11	0.42±0.12	0.63±0.10	0.73±0.05
F值	0.873	6.170	5.087	19.958	35.958
P值	0.438	0.011	0.021	<0.001	<0.001

组别	迁移细胞数/ （个/高倍镜视野）	侵袭细胞数/ （个/高倍镜视野）
sh-MUC16组	108.83±7.57	93.50±9.29
sh-Control组	131.33±11.20^*	122.25±7.78^*
空白组	127.83±9.22^*	127.00±9.92^*
F值	9.853	26.034
P值	0.002	<0.001

组别	迁移细胞数/ （个/高倍镜视野）	侵袭细胞数/ （个/高倍镜视野）
sh-MUC16组	108.83±7.57	93.50±9.29
sh-Control组	131.33±11.20^*	122.25±7.78^*
空白组	127.83±9.22^*	127.00±9.92^*
F值	9.853	26.034
P值	0.002	<0.001

组别	MUC16	E-Cad	N-Cad
sh-MUC16组	0.21±0.05	0.65±0.05	0.27±0.12
sh-Control组	0.72±0.11^*	0.23±0.09^*	0.86±0.05^*
空白组	0.76±0.16^*	0.28±0.04^*	0.79±0.04^*
F值	42.627	76.559	105.144
P值	<0.001	<0.001	<0.001