藤梨根提取物对口腔鳞状细胞癌细胞凋亡、侵袭及迁移的影响及机制研究  

Effect and Mechanism of Extract from Actinidia Chinensis Radix on Apoptosis,Invasion and Migration of Oral Squamous Cell Carcinoma Cells

作　　者：赵愧云 袁慧芳[1] ZHAO Kuiyun;YUAN Huifang

机构地区：[1]河南中医药大学第三附属医院口腔科,河南郑州450004

出　　处：《新中医》2022年第3期164-170,共7页New Chinese Medicine

摘　　要：目的:探讨藤梨根提取物对口腔鳞状细胞癌细胞凋亡、侵袭及迁移的影响及分子机制。方法:将口腔鳞状细胞癌CAL-27细胞分为对照组,藤梨根低、中、高剂量组,藤梨根高剂量+pcDNA组,藤梨根高剂量+pcDNA-SBF2-AS1组。流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;蛋白质印迹(Western blot)法检测E钙黏蛋白(E-cadherin)、N钙黏蛋白(N-cadherin)、裂解天冬氨酸特异性半胱氨酸蛋白酶3 (Cleaved-caspase3)、前体caspase3 (Pro-caspase3)蛋白表达;实时荧光定量PCR (RT-qPCR)检测SET结合因子2反义RNA1 (SBF2-AS1)和miR-329的表达水平;双荧光素酶报告实验验证SBF2-AS1和miR-329的靶向关系。结果:不同剂量藤梨根提取物处理后,口腔鳞状细胞癌CAL-27细胞中细胞凋亡率升高,迁移侵袭细胞数降低,E-cadherin、Cleaved-caspase3表达水平升高,N-cadherin、Pro-caspase3表达水平降低,SBF2-AS1表达水平降低,miR-329表达水平升高,且呈剂量依赖性(P<0.05)。过表达SBF2-AS1可逆转藤梨根提取物对CAL-27凋亡、迁移侵袭的影响。SBF2-AS1靶向miR-329。结论:藤梨根提取物可能通过调控SBF2-AS1/miR-329抑制口腔鳞状细胞癌细胞侵袭及迁移,促进细胞凋亡。Objective: To investigate the effect and molecular mechanism of Actinidia chinensis radix extract on apoptosis,invasion and migration of oral squamous cell carcinoma cells. Methods:Oral squamous cell carcinoma CAL-27 cells were divided into control group, low, medium and high dose Actinidia chinensis radix groups, high dose Actinidia chinensis radix+ pcDNA group and high dose Actinidia chinensis radix+ pcdna-SBF2-AS1 group. Apoptosis was detected by flow cytometry.Transwell detected cell migration and invasion. Western blot was used to detect the protein expression of E-cadherin, N-cadherin, Cleaved cysteinyl aspartate specific proteinase3(Cleaved-caspase3) and Pro-caspase3. The expression levels of set binding factor 2 antisense RNA1(SBF2-AS1) and miR-329 were detected by real-time fluorescence quantitative PCR(RT-qPCR). Double Luciferase Report experiment verified the targeting relationship between SBF2-AS1 and miR-329. Results: After treatment with different doses of Actinidia chinensis radix extract, the apoptosis rate of oral squamous cell CAL-27 increased, the number of migrating and invasive cells decreased, the expression levels of Ecadherin and Cleaved-caspase3 increased, the expression levels of N-cadherin and Pro-caspase3 decreased, the expression level of SBF2-AS1 decreased,and the expression level of miR-329 increased in a dose-dependent manner(P<0.05). Overexpression of SBF2-AS1 could reverse the effects of Actinidia chinensis radix extract on apoptosis,migration and invasion of CAL-27. SBF2-AS1 targets mi R-329. Conclusion:Actinidia chinensis radix extract may inhibit the invasion and migration of oral squamous cell carcinoma cells and promote apoptosis by regulating SBF2-AS1/miR-329.

关 键 词：口腔鳞状细胞癌 藤梨根提取物 SBF2-AS1 miR-329 凋亡 侵袭 迁移 

分 类 号：R739.85[医药卫生—肿瘤]