[RNA干扰人滋养层细胞表面抗原2基因表达对舌鳞状细胞癌增殖与凋亡的影响及其机制]
[Effect of human trophoblast cell-surface antigen 2 gene expression by RNA interference on proliferation and apoptosis of tongue squamous cell carcinoma and its mechanism].
作者信息
Fang Z, Chen S, Zhao J F, Sun Q, Qiu F, Li X M
机构信息
Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
Department of Pathology, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
出版信息
Zhonghua Kou Qiang Yi Xue Za Zhi. 2018 Sep 9;53(9):640-644. doi: 10.3760/cma.j.issn.1002-0098.2018.09.015.
To investigate the effect and its mechanism of human trophoblast cell-surface antigen 2 (Trop2) gene expression was inhibited in squamous cell carcinoma of tongue on the proliferation and apoptosis of cancer cells. A total of 46 patients from February 2014 to May 2016 received radical treatment of tongue cancer from oral and maxillofacial surgery of the First Affiliated Hospital of Zhengzhou University were enrolled in this study. Real time PCR and Western blotting were used to detect mRNA and protein expression of Trop2 in tongue squamous cell carcinoma and corresponding adjacent tissues; NC-siRNA and Trop2-siRNA were transfected into human tongue squamous cell carcinoma CAL-27 cells, a blank control group (control) was set, the expression of Trop2, Ki-67, cyclin D1, cleaved caspase3, Notch1, Hes1 protein after transfected for 48 h were detected by Western bloting; cell proliferation was detected by cell counting kit-8; cell cycle and apoptosis rate were detected by flow cytometry. The mRNA (5.72±1.13) and protein expression (0.77±0.06) of Trop2 gene in tongue squamous cell carcinoma were significantly higher than those in adjacent tissues (0.92±0.15, 0.11±0.01, 0.05); Trop2 protein expression after transfeced Trop2-siRNA (0.08±0.01) in CAL-27 cells decreased significantly (0.46±0.05); the survival rate (64.28±4.12)%, S cells (14.54±1.02)% and Ki-67 (0.12±0.01), cyclin D1 (0.04±0.01) protein expression in Trop2-siRNA group were significantly lower than those in control group [(100.00±1.02)%, (27.33±1.11)%, (0.24±0.02), (0.20±0.02)] and NC-siRNA group [(96.55±2.43)%, (26.67±1.23)%, (0.26±0.03), (0.21±0.024)], the apoptosis rate (23.55±1.45)%, G0/G1 cells (72.32±0.94)% and the expression of cleaved caspase 3 (0.16±0.02), Notch1 (0.62±0.06) and Hes1 (0.50±0.05) protein were significantly higher than control group and NC-siRNA group (0.05). The expression of Trop2 gene was inhibited in tongue squamous carcinoma cells can reduce the proliferation of cancer cells, block the cells in phase G1, and promote the apoptosis of cells. The mechanism is related to the up regulation of Notch1 signaling pathway.
探讨抑制人滋养层细胞表面抗原2（Trop2）基因表达对舌鳞状细胞癌癌细胞增殖和凋亡的影响及其机制。选取2014年2月至2016年5月在郑州大学第一附属医院口腔颌面外科接受舌癌根治术的46例患者纳入本研究。采用实时荧光定量PCR和蛋白质免疫印迹法检测Trop2在舌鳞状细胞癌及相应癌旁组织中的mRNA和蛋白表达；将NC-siRNA和Trop2-siRNA转染入人舌鳞状细胞癌CAL-27细胞，设空白对照组（control），转染48 h后采用蛋白质免疫印迹法检测Trop2、Ki-67、细胞周期蛋白D1、裂解的半胱天冬酶3、Notch1、Hes1蛋白的表达；采用细胞计数试剂盒-8检测细胞增殖；采用流式细胞术检测细胞周期和凋亡率。舌鳞状细胞癌中Trop2基因的mRNA（5.72±1.13）和蛋白表达（0.77±0.06）显著高于癌旁组织（0.92±0.15，0.11±0.01，0.05）；CAL-27细胞转染Trop2-siRNA后Trop2蛋白表达（0.08±0.01）较对照组（0.46±0.05）显著降低；Trop2-siRNA组的生存率（64.28±4.12）%、S期细胞（14.54±1.02）%以及Ki-67（0.12±0.01）、细胞周期蛋白D1（0.04±0.01）蛋白表达均显著低于对照组[（100.00±1.02）%，（27.33±1.11）%，（0.24±0.02），（0.20±0.02）]和NC-siRNA组[（96.55±2.43）%，（26.67±1.23）%，（0.26±0.03），（0.21±0.024）]，凋亡率（23.55±1.45）%、G0/G1期细胞（72.32±0.94）%以及裂解的半胱天冬酶3（0.16±0.02）、Notch1（0.62±0.06）和Hes1（0.50±0.05）蛋白表达均显著高于对照组和NC-siRNA组（P<0.05）。抑制舌鳞状癌细胞中Trop2基因的表达可降低癌细胞的增殖，使细胞阻滞于G1期，并促进细胞凋亡。其机制与Notch1信号通路的上调有关。
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