辛伐他汀对唾液腺腺样囊性癌细胞生长及survivin表达的影响

摘要/Abstract

摘要： 目的探讨辛伐他汀（simvastatin）对人唾液腺腺样囊性癌细胞株SACC-83细胞增殖、凋亡及survivin蛋白表达的影响。方法体外培养SACC-83细胞,用不同浓度的辛伐他汀（0、10、20、30、40、50 μmol/L）对体外培养的SACC-83细胞进行不同时间的干预,CCK-8法检测药物对细胞增殖能力的影响;结晶紫染色观察辛伐他汀对细胞集落形成的影响;Annexin-V/PI双染色流式细胞术检测不同浓度辛伐他汀作用于SACC-83细胞48 h后凋亡细胞的百分率;Western蛋白印迹检测 survivin蛋白表达的变化。采用SPSS13.0软件包对数据进行单因素方差分析和q检验。结果用不同浓度的辛伐他汀处理细胞不同时间后,SACC-83的生长受到明显抑制,并具有时间、浓度梯度依赖性;结晶紫染色观察到细胞集落形成也受到抑制（P<0.05）;药物处理48 h后,细胞凋亡率随药物浓度增加而升高（从8.60% 到67.97%）;同时,SACC-83细胞survivin蛋白的表达量显著降低（P<0.05）。结论辛伐他汀能抑制SACC-83细胞的增殖,诱导细胞凋亡,其机制可能与survivin蛋白的表达下调有关。

关键词: 辛伐他汀, 唾液腺腺样囊性癌, 增殖, 凋亡, Survivin

Abstract: PURPOSE: To investigate the effect of simvastatin on proliferation, apoptosis and expression of survivin in salivary adenoid cystic carcinoma SACC-83 cells. METHODS: SACC-83 cells were cultured in vitro and exposed to different concentrations of simvastatin (0, 10, 20, 30, 40, 50 μmol/L) for 24h and 48h, respectively. Cell survival rate was calculated with CCK-8 assay and the OD values were calculated. Colony formation of SACC-83 cells was observed following a 10-day of exposure to different concentrations of simvastatin. The percentage of apoptotic cells was determined by flow cytometry following annexin-V/Pl staining. At the same time, expression of survivin protein was quantitatively analyzed by Western blotting. The data were analyzed by one-way ANOVA and repeated measurement design using SPSS 13.0 software package. RESULTS: At the concentration of 0, 10, 20, 30, 40, 50 μmol/L, CCK-8 method showed that simvastatin could inhibit the proliferation of SACC-83 cells in a dose- and time-dependent manner. After treatment with simvastatin for 48 h, the IC50 value was 26.27 μmol/L. Simvastatin significantly suppressed the colonization of SACC-83 cell in a dose-dependent manner. Flow cytometry indicated that simvastatin over 10μmol/L increased the cell populations of both the early apoptotic and late apoptotic cells in a dose dependent manner. Simvastatin at 10, 30 and 50 μmol/L resulted in apoptotic percentages of (21.43±5.43)%, (42.8±10.08)%, and (67.97±12.5)%, respectively at 48h of exposure. In addition, simvastatin down-regulated survivin in SACC-83 cells whereas survivin was over-expressed in the untreated SACC-83 cells. The difference was significant (P<0.05). CONCLUSIONS: Simvastatin can significantly inhibit proliferation of SACC-83 cells and induce apoptosis, as indicated by lower-expression of survivin, which suggests that survivin might serve as a novel target in the therapy for salivary adenoid cystic carcinoma.

Key words: Simvastatin, Salivary adenoid cystic carcinoma, Proliferation, Apoptosis, Survivin

中图分类号:

R739.8

蔡文燕, 严斐, 董晶, 孙晋虎. 辛伐他汀对唾液腺腺样囊性癌细胞生长及survivin表达的影响[J]. 中国口腔颌面外科杂志, 2016, 14(3): 213-217.

CAI Wen-yan, YAN Fei, DONG Jing, SUN Jin-hu. Effect of simvastatin on proliferation and apoptosis of salivary adenoid cystic carcinoma SACC-83 cells and survivin expression[J]. China J Oral Maxillofac Surg, 2016, 14(3): 213-217.

参考文献

[1] 热孜亚·艾尼, 迪丽努尔·阿西木, 孙振柱, 等. 涎腺腺样囊性癌的局部复发及远处转移因素分析 [J]. 实用口腔医学杂志, 2011, 27(5): 649-652.
[2] Schoenfeld JD, Sher DJ, Norris CM Jr, et al. Salivary gland tumors treated with adjuvant intensity-modulated radiotherapy with or without concurrent chemotherapy [J]. Int J Radiat Oncol Biol Phys, 2012, 82(1): 308-314.
[3] Shen C, Xu T, Huang C, et al. Treatment outcomes and prognostic features in adenoid cystic carcinoma originated from the head and neck [J]. Oral Oncol, 2012, 48(5): 445-449.
[4] 贾志宇, 邸永宾, 李晓玲, 等. 吴茱萸碱抑制唾液腺腺样囊性癌细胞系ACC-M增殖的实验研究 [J]. 实用口腔医学杂志, 2012, 28(5): 592-596.
[5] Borahay MA, Kilic GS, Yallampalli C, et al. Simvastatin potently induces calcium-dependent apoptosis of human leiomyoma cells [J]. J Biol Chem, 2014, 289(51): 35075-35086.
[6] Lee KJ, Moon JY, Choi HK, et al. Immune regulatory effects of simvastatin on regulatory T cell-mediated tumour immune tolerance [J]. Clin Exp Immunol, 2010, 161(2): 298-305.
[7] 刘蔚, 张连峰, 张宇恒. 辛伐他汀抑制HepG2细胞增殖作用的机制 [J]. 中华肝脏病杂志, 2010, 18(10): 751-753.
[8] Chang HL, Chen CY, Hsu YF, et al. Simvastatin induced HCT116 colorectal cancer cell apoptosis through p38MAPK-p53-survivin signaling cascade[J]. Biochim Biophys Acta, 2013, 1830(8): 4053-4064.
[9] Otsuki T, Kanno T, Fujita Y, et al. A3 adenosine receptor-mediated p53-dependent apoptosis in Lu-65 human lung cancer cells [J]. Cell Physiol Biochem, 2012, 30(1): 210-220.
[10] 贾志宇, 庄志征, 岳磊, 等. 莱菔硫烷对涎腺腺样囊性癌细胞系ACC-M增殖和凋亡的影响[J]. 实用口腔医学杂志, 2014, 30(5): 653-657.
[11] 王柏欣, 陈梅, 王淑秋, 等. 辛伐他汀对大鼠脑缺血再灌注损伤后线粒体游离钙, 细胞色素C表达水平的影响[J]. 中国老年医学杂志, 2011, 31(19): 3764-3765.
[12] Shankar SL, Mani S, O'guin KN, et al. Survivin inhibition induces human neural tumor cell death through caspase-independent and -dependent pathways[J]. J Neurochem, 2001, 79(2): 426-436.
[13] Wang Y, Xu SL, Wu YZ, et al. Simvastatin induces caspase-dependent apoptosis and activates P53 in OCM-1 cells[J]. Exp Eye Res, 2013, 113: 128-134.
[14] Chan KK, Oza AM, Siu LL. The statins as anticancer agents [J]. Clin Cancer Res, 2003, 9(1): 10-19.
[15] 杨士杰, 姜达. 辛伐他汀对SMMC-7721的凋亡作用及机制研究[J]. 河北医药, 2012, 34(4): 591-592.
[16] 米磊, 张斌, 王岩松, 等. 熊果酸对ACC-83增殖抑制和诱导凋亡的观察[J]. 口腔颌面外科杂志, 2010, 20(6): 389-393.
[17] Mitsiades CS, Mitsiades N, Poulaki V, et al. Activation of NF-kappaB and upregulation of intracellular anti-apoptotic proteins via the IGF-1/Akt signaling in human multiple myeloma cells: therapeutic implications [J]. Oncogene, 2002, 21(37): 5673-5683.
[18] Jaiswal PK, Goel A, Mittal RD. Survivin: a molecular biomarker in cancer [J]. Indian J Med Internet Res, 2015, 141(4): 389-397.
[19] Waligorska-Stachura J, Jankowska A, Wasko R, et al. Survivin-prognostic tumor biomarker in human neoplasms-review[J]. Ginekol Pol, 2012, 83(7): 537-540.
[20] 肖胤, 杨军, 肖林, 等. Survivin基因在人唾液腺腺样囊性癌中的表达 [J]. 临床口腔医学杂志, 2007, 23(4): 209-211.
[21] 张斌, 牟海滨, 徐旭光, 等. 生存素基因在三氧化二砷诱导腺样囊性癌-2细胞凋亡中的作用 [J]. 华西口腔医学杂志, 2010, 28(3): 246-249.