白藜芦醇对肺癌细胞95D增殖和侵袭的影响

doi:10.3760/cma.j.issn.1673-422X.2018.12.001

摘要/Abstract

摘要： 目的研究白藜芦醇对人肺腺癌95D细胞株增殖及侵袭能力的影响。方法采用0、10、20、40、80 mol/L白藜芦醇处理人肺腺癌95D细胞，以0 mol/L组作为对照。四甲基偶氮唑蓝（MTT）法测定95D细胞增殖水平；流式细胞仪检测细胞周期和细胞凋亡；体外黏附实验测定细胞黏附率；Transwell小室测定细胞侵袭力；荧光免疫细胞化学方法测定基质金属蛋白酶2(MMP2)和金属蛋白酶组织抑制剂2(TIMP2)的表达。结果白藜芦醇作用72 h，0、10、20、40、80 mol/L白藜芦醇处理组细胞增殖率分别为100%、（82.23±0.33）%、（62.45±0.27）%、（49.89±0.43）%、（45.11±0.35）%，组间差异有统计学意义（F=87.830，P=0.002）；10、20、40、80 μmol/L组细胞增殖率均较对照组降低（P=0.017，P<0.001，P<0.001，P<0.001）。白藜芦醇作用48 h，各组细胞凋亡率分别为0、（34.90±0.91）%、（41.33±0.13）%、（45.47±0.87）%、（59.46±0.59）%，组间差异有统计学意义（F=21.032，P=0.002）；10、20、40、80 mol/L组细胞凋亡率均较对照组增加（P=0.001，P<0.001，P<0.001，P<0.001）。白藜芦醇作用48 h，各组S期细胞比例分别为（18.12±0.62）%、（38.33±0.62）%、（54.15±0.74）%、（44.85±0.82）%、（50.01±0.35）%，组间差异有统计学意义（F=104.156，P=0.001）；10、20、40、80 μmol/L组S期细胞比例均较对照组增加（P=0.001，P<0.001，P<0.001，P<0.001）。白藜芦醇作用24 h，各组细胞黏附率分别为100%、（87.41±0.02）%、（84.32±0.03）%、（68.23±0.04）%、（63.01±0.02）%，组间差异有统计学意义（F=13.760，P<0.001）；20、40、80 μmol/L组细胞黏附率均较对照组降低（P=0.035，P<0.001，P<0.001），而10 mol/L组与对照组差异无统计学意义（P=0.058）。白藜芦醇作用24 h，各组细胞侵袭率分别为100%、（97.01±0.03）%、（74.89±0.07）%、（34.07±0.03）%、（14.65±0.02）%，组间差异有统计学意义（F=39.382，P=0.001）；20、40、80 μmol/L组细胞侵袭率均较对照组降低（P=0.012，P<0.001，P<0.001），而10 μmol/L组与对照组差异无统计学意义（P=0.881）。白藜芦醇作用48 h，与对照组相比，20 mol/L组MMP2蛋白表达量降低，TIMP2蛋白表达量增加。结论白藜芦醇可抑制人肺腺癌95D细胞增殖，并具有抗肿瘤细胞侵袭作用，其机制可能涉及对MMP2/TIMP2表达的双向调控。

关键词: 肺肿瘤, 细胞增殖, 细胞黏附, 肿瘤侵润, 白藜芦醇

Abstract: ObjectiveTo study the effects of resveratrol on the proliferation and invasion capacity of human lung adenocarcinoma 95D cells. MethodsHuman lung adenocarcinoma 95D cells were treated with 0, 10, 20, 40, 80 μmol/L resveratrol and treated with 0 μmol/L as the control group. The proliferation level of 95D cells was measured by methyl thiazolyl tetrazolium (MTT). Cell cycle and apoptosis were detected by flow cytometry. Cell adhesion rate was determined by in vitro adhesion test. Cell invasiveness was measured by Transwell chamber. The expressions of matrix metalloproteinase2 (MMP2) and tissue inhibitior of metalloproteinase2 (TIMP2) were determined by fluorescent immunocytochemistry. ResultsWhen the 95D cells were treated with resveratrol for 72 h, the cell proliferation rates in groups treated with 0, 10, 20, 40, 80 μmol/L resveratrol were 100%, (82.23±0.33)%, (62.45±0.27)%, (49.89±0.43)%, (45.11±0.35)% respectively, with a significant difference (F=87.830, P=0.002). The proliferation rates of 95D cells in the 10, 20, 40, 80 μmol/L resveratrol groups were significantly inhibited compared with the control group (P=0.017, P<0.001, P<0.001, P<0.001). When the 95D cells were treated for 48 h, the apoptosis rates of cells in each group were 0, (34.90±0.91)%, (41.33±0.13)%, (45.47±0.87)%, (59.46±0.59)% respectively, with a significant difference (F=21.032, P=0.002). The apoptosis rates of 95D cells in the 10, 20, 40, 80 μmol/L resveratrol groups were significantly increased compared with the control group (P=0.001, P<0.001, P<0.001, P<0.001). When the 95D cells were treated for 48 h, the S phase cell percentages in each group were (18.12±0.62)%, (38.33±0.62)%, (54.15±0.74)%, (44.85±0.82)%, (50.01±0.35)% respectively, with a significant difference (F=104.156, P=0.001). The S phase cell percentages in the 10, 20, 40, 80 μmol/L resveratrol groups were significantly higher compared with the control group (P=0.001, P<0.001, P<0.001, P<0.001). When the 95D cells were treated for 24 h, the cell adhesion rates in each group were 100%, (87.41±0.02)%, (84.32±0.03)%, (68.23±0.04)%, (63.01±0.02)% respectively, with a significant difference (F=13.760, P<0.001). The cell adhesion rates in the 20, 40, 80 μmol/L resveratrol groups were significantly inhibited compared with the control group (P=0.035, P<0.001, P<0.001), while the 10 μmol/L resveratrol group had no significant difference compared with the control group (P=0.058). When the 95D cells were treated for 24 h, the cell invasion rates in each group were 100%, (97.01±0.03)%, (74.89±0.07)%, (34.07±0.03)%, (14.65±0.02)% respectively, with a significant difference (F=39.382, P=0.001). The cell invasion rates in the 20, 40, 80 μmol/L groups were significantly inhibited compared with the control group (P=0.012, P<0.001, P<0.001), while the 10 μmol/L resveratrol group had no significant difference compared with the control group (P=0.881). When the 95D cells were treated for 48 h, MMP2 protein expression was decreased in the 20 mol/L group, while TIMP2 protein expression was increased compared with the control group. ConclusionResveratrol can inhibit the proliferation of human lung adenocarcinoma 95D cells and has effect of antitumor cell invasiveness, and its mechanism may involve twoway regulation of MMP2/TIMP2 expression.

Key words: Lung neoplasms, Cell proliferation, Cell adhesion, Neoplasm invasiveness;, Resveratrol

陈海霞. 白藜芦醇对肺癌细胞95D增殖和侵袭的影响[J]. 国际肿瘤学杂志, 2018, 45(12): 705-710.

Chen Haixia. Effects of resveratrol on proliferation and invasion of lung cancer 95D cells[J]. Journal of International Oncology, 2018, 45(12): 705-710.

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