Effect of Calpain 1 Gene Silencing by RNA Interference on Proliferation and Migration of Oral Squamous Cell Carcinoma Cell Lines HSC3

doi:10.13701/j.cnki.kqyxyj.2018.02.011

Abstract

Abstract: Objective: To explore the silencing effect of calpain 1 by RNA interference on cell proliferation and migration in oral squamous cell carcinoma cell line HSC3. Methods: siRNA expression vector (siRNA-calpain 1) against the calpain 1 gene was transiently transfected into HSC3. The proliferation was tested by MTT method. The cell cycle and apoptosis was detected by flow cytometry. The cell migration and invasion were assessed by Transwell assay. The expression of TGF-β RII, Vimentin, Smad2, Smad3, phosphorylated Smad2/3, E-cadherin, N-cadherin, MMP-2, and MMP-9 protein were detected by western blot. Results: The expression of calpain 1 in HSC3 cell lines was significantly higher than those in NOKSI cell lines (P<0.05). Compared with the blank group and the si-NC group, the expression of calpain 1 was significantly decreased (P<0.05). Compared with si-NC group, the viability of si-calpain group was significantly decreased after 48 h, 76 h and 96 h after transfection with si-calpain1 (P<0.05). The proportion of G0/G1 phase cells in si-calpain group was significantly increased, while the proportion of S phase cells was significantly decreased (P<0.05). The expression of E-cadherin in si-calpain group was significantly increased, while cyclin-D1, cyclin-E, Vimentin, N-cadherin, MMP-2, MMP-9, TGF-β RII, Smad2, Smad3, p-Smad2/3 protein expression were significantly decreased (P<0.05). Conclusion: siRNA silencing calpain 1 gene can inhibit cell proliferation by inhibiting cell cycle G0/G1, and inhibit cell migration and invasion by inhibiting MMPs and EMT. This process involves TGF-β/Smad signaling pathway.

Key words: Oral squamous cell carcinoma, Calpain 1, Cell line HSC3, Cell proliferation, Cell migration

CLC Number:

R739.8

ZHANG Hou-hua, FAN Hua-li. Effect of Calpain 1 Gene Silencing by RNA Interference on Proliferation and Migration of Oral Squamous Cell Carcinoma Cell Lines HSC3[J]. Journal of Oral Science Research, 2018, 34(2): 152-156.

References

[1] 刘盼,多力昆·吾甫尔.鳞状细胞癌抗原在口腔鳞癌诊治中的应用[J]. 现代口腔医学杂志,2017,31(1)∶49-52
[2] 夏冬景,黄莹莹,李海朋,等.磷酸二酯酶4D在口腔鳞状细胞癌发生发展过程中的作用研究[J]. 口腔医学研究,2017,33(6)∶663-667
[3] 安静,朱筑霞,王红梅,等.钙蛋白酶抑制EGF诱导雌激素受体阴性乳腺癌细胞体外增殖[J]. 基础医学与临床,2015,35(3)∶336-340
[4] Liu B, Zhou Y, Lu D, et al. Comparison of the protein expression of calpain-1, calpain-2, calpastatin and calmodulin between gastric cancer and normal gastric mucosa[J]. Oncology Letters, 2017, 14(3)∶3705-3710
[5] 曾伟,南清振,戴益琛,等.大肠腺瘤与大肠腺癌肿瘤相关基因的差异表达[J]. 世界华人消化杂志,2011(8)∶868-873
[6] Storr S J, Pu X, Davis J, et al. Expression of the calpain system is associated with poor clinical outcome in gastro-oesophageal adenocarcinomas.[J]. Journal of Gastroenterology, 2013, 48(11)∶1213-1221
[7] 陈华妹,李铮,王旭东,等.钙蛋白酶-周期蛋白E介导EGF诱导的MCF-7乳腺癌细胞迁移能力增强[J]. 贵阳医学院学报,2015,40(12)∶1344-1347
[8] Santra MK, Narendra W, Green MR. F-Box Protein FBXO31 Mediates Cyclin D1 Degradation to Induce G1 Arrest Following DNA Damage[J]. Nature, 2009, 459(7247)∶722-725
[9] 艾军华,时军.肿瘤干细胞和上皮-间质转化之间的关系及其在肿瘤侵袭转移中的作用[J]. 医学综述,2015,21(12)∶2180-2183
[10] 祝丽晶,李芹芹,刘强,等.血清基质金属蛋白酶-9水平在非小细胞肺癌侵袭和转移中意义[J]. 中华实用诊断与治疗杂志,2017,31(2)∶165-166
[11] 焦慧,高志光,高发鹏,等.基质金属蛋白酶-2沉默对喉癌侵袭生长的影响[J]. 系统医学,2017,2(1)∶10-12
[12] 汪婷,程真顺,刘媛,等.TGF-β1/Smad2通路参与IL-17诱导的A549细胞上皮-间质转化[J]. 武汉大学学报(医学版),2017,38(5)∶719-723
[13] 邢洲,张惠,胡慧贤,等.CCL18与 TGF-β促进乳腺癌 EMT 的比较[J]. 暨南大学学报(自然科学与医学版),2016,37(5)∶367-371
[14] Tan WJ, Tan QY, Wang T, et al. Calpain 1 regulates TGF-尾1-induced epithelial-mesenchymal transition in human lung epithelial cells via PI3K/Akt signaling pathway[J]. Am J Transl Res, 2017, 9(3)∶1402-1409

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