Effects of Silencing RohA Gene on Proliferation and Invasion of Salivary Adenoid Cystic Carcinoma Cell Line ACC-2

doi:10.3969/j.issn.1005-4979.2018.01.005

Abstract

Abstract: Objective: To investigate the effects of silencing RohA gene on proliferation and invasion of salivary adenoid cystic carcinoma cell line ACC-2. Methods: ACC-2 cells were cultured and were divided into siRNA-RhoA group, siRNA-control sequence group and blank control group. Real-time fluorescence quantitative PCR was used to detect the expression of Rho gene. The expression of RohA protein was detected by Western blot. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect the apoptosis of each group. Cell migration and invasion was detected by using Transwell method. Results: Compared with siRNA-control sequence group and blank control group, the relative expression level of RohA mRNA and protein in siRNA-RhoA group were decreased, the difference were statistically significant (P<0.05). Compared with siRNA-control sequence group and blank control group, the absorbance A values at 24 h, 48 h, 72 h and 96 h in the siRNA-RhoA group were decreased, the differences were statistically significant (P<0.05). Compared with siRNA-control sequence group and blank control group, the cell apoptosis rate in the siRNA-RhoA group was significantly increased, the difference was statistically significant (P<0.05). Compared with siRNA-control sequence group and blank control group, the number of migrating cells and the number of invasive cells in the siRNA-RhoA group were decreased, the differences were statistically significant(P<0.05). Conclusion: Specific silencing of RohA gene expression could significantly reduce ACC-2 proliferation, promote apoptosis, inhibit cell metastasis and invasion.

Key words: salivary adenoid cystic carcinoma, RohA, small interference RNA, cell proliferation, cell invasion

摘要： 目的：探讨沉默RohA基因对涎腺腺样囊性癌（SACC）细胞株ACC-2增殖和侵袭的影响。方法：培养ACC-2细胞，分为siRNA-RhoA组、siRNA-对照序列组和空白对照组，实时荧光定量PCR技术检测细胞中Rho基因表达，Western blot法检测细胞中RohA蛋白表达，MTT法检测细胞增殖能力，流式细胞术检测各组细胞凋亡，Transwell法检测细胞迁移和侵袭能力。结果：与siRNA-对照序列组和空白对照组比较，siRNA-RhoA组细胞中RohA mRNA和蛋白相对表达量均降低，差异均有统计学意义（P<0.05）；与siRNA-对照序列组和空白对照组比较，siRNA-RhoA组细胞在24、48、72、96h时吸光度A值均降低，差异有统计学意义（P<0.05）；与siRNA-对照序列组和空白对照组比较，siRNA-RhoA组细胞凋亡率明显增加，差异有统计学意义（P<0.05）；与siRNA-对照序列组和空白对照组比较，siRNA-RhoA组迁移细胞数和侵袭细胞数均减少，差异均有统计学意义（P<0.05）。结论：特异性沉默RohA基因表达可明显减弱ACC-2增殖能力，促进细胞凋亡，抑制细胞转移和侵袭能力。

关键词: 涎腺腺样囊性癌, RohA, 小分子RNA干扰, 细胞增殖, 细胞侵袭

CLC Number:

R782
739.8

WANG Xiao-long, CAO Shun-shun, SHU Chuan-ji. Effects of Silencing RohA Gene on Proliferation and Invasion of Salivary Adenoid Cystic Carcinoma Cell Line ACC-2[J]. 《Journal of Oral and Maxillofacial Surgery》, doi: 10.3969/j.issn.1005-4979.2018.01.005.

汪晓龙，曹顺顺，舒传继. 沉默RohA基因对涎腺腺样囊性癌细胞株ACC-2增殖和侵袭的影响[J]. 《口腔颌面外科杂志》, doi: 10.3969/j.issn.1005-4979.2018.01.005.

/ / Recommend / Download Citations

URL: https://journal06.magtech.org.cn/Jweb_joms/EN/10.3969/j.issn.1005-4979.2018.01.005

https://journal06.magtech.org.cn/Jweb_joms/EN/Y2018/V28/I1/26

References

[1] Li Q, Huang P, Zheng C, et al. Prognostic significance of p53 immunohistochemical expression in adenoid cystic carcinoma of the salivary glands: a meta-analysis[J]. Oncotarget, 2017, 8(17):29458-29473.

[2] Dantas AN, Morais EF, Macedo RA, et al. Clinicopathological characteristics and perineural invasion in adenoid cystic carcinoma: a systematic review [J]. Braz J Otorhinolaryngol, 2015, 81(3):329-335.

[3] Otsu K, Harada H. Rho GTPases in ameloblast differentiation[J]. Jpn Dent Sci Rev, 2016, 52(2):32-40.

[4] Hetmanski JH, Schwartz JM, Caswell PT. Modelling GTPase dynamics to understand RhoA-driven cancer cell invasion[J]. Biochem Soc Trans, 2016, 44(6):1695-1700.

[5] Cai Q, Zhang R, Wu G, et al. Adenoid cystic carcinoma of submandibular salivary gland with late metastases to lung and choroid: a case report and literature review[J]. J Oral Maxillofac Surg, 2014, 72(9):1744-1755.

[6] Wysocki PT, Izumchenko E, Meir J, et al. Adenoid cystic carcinoma: emerging role of translocations and gene fusions[J]. Oncotarget, 2016, 7(40):66239-66254.

[7] Watzlawick R, Sena ES, Dirnagl U, et al. Effect and reporting bias of RhoA/ROCK-blockade intervention on locomotor recovery after spinal cord injury: a systematic review and meta-analysis[J]. JAMA Neurol, 2014, 71(1):91-99.

[8] Jeong D, Park S, Kim H, et al. RhoA is associated with invasion and poor prognosis in colorectal cancer[J]. Int J Oncol, 2016, 48(2):714-722.

[9] 廖山婴, 刘超, 王蓓蓓, 等. RhoA/ROCK信号通路在胃癌中的表达及其与胃癌生物学行为的关系[J]. 胃肠病学和肝病学杂志, 2017, 26(3):251-255.

[10] 胡威, 董忠谊, 吴德华. FAT10通过激活RhoA介导肝细胞性肝癌侵袭转移[J]. 中国肿瘤临床, 2015, 42(14):689-694.

[11] Timpson P, McGhee EJ, Morton JP, et al. Spatial regulation of RhoA activity during pancreatic cancer cell invasion driven by mutant p53[J]. Cancer Res, 2011, 71(3):747-757.

[12] 严国鑫, 樊兵, 邹荣海, 等. 沉默RohA基因对舌鳞状细胞癌细胞增殖的影响[J]. 华西口腔医学杂志, 2016, 34(6):620-625.

Please choose a citation manager

Content to export