姜黄素联合顺铂对HepG2肝癌细胞增殖、凋亡、迁移及侵袭的影响

Effects of curcumin combined with cisplatin on cell proliferation, apoptosis, migration and invasion in hepatocarcinoma HepG2 cell

目的观察姜黄素联合顺铂对HepG2肝癌细胞的增殖、凋亡、迁移及侵袭的影响。方法肝癌HepG2细胞加药处理，分为二甲基亚砜（DMSO）对照组、姜黄素处理组（5、10、20 μmol/L）、顺铂处理组（1、5、10 μmol/L）和联合用药处理组（5 μmol/L姜黄素＋5 μmol/L顺铂、10 μmol/L姜黄素＋5 μmol/L顺铂）。使用噻唑蓝（MTT）法检测不同药物处理后HepG2细胞的增殖能力的变化；使用Hoechst 33258检测药物处理后细胞凋亡的变化；应用划痕愈合实验和Transwell侵袭实验检测药物处理后肿瘤细胞的迁移和侵袭能力的变化。结果MTT结果显示联合用药组与5 μmol/L顺铂组比较细胞存活率明显降低[（40.27±4.80）%比（61.80±9.14）%，P=0.023；（19.37±5.02）%比（61.80±9.14）%，P=0.002]；Hoechst 33258染核10 μmol/L姜黄素＋5 μmol/L顺铂组与5 μmol/L顺铂组比较细胞凋亡率增加[（18.95±1.30）%比（10.35±1.46）%，P=0.012]；划痕愈合实验结果显示联合用药组较5 μmol/L顺铂组迁移愈合率明显下降[（33.81±1.63）%比（39.77±2.48）%，P=0.026；（24.54±4.93）%比（39.77±2.48）%，P=0.003]，联合用药后可抑制细胞迁移能力；Transwell侵袭实验示联合用药组侵袭细胞数较5 μmol/L顺铂组比较明显减少[（24.33±2.52）个比（32.00±3.61）个，P=0.039；（15.67±3.06）个比（32.00±3.61）个，P=0.004]，提示联合用药后细胞侵袭能力下降。结论姜黄素联合顺铂可以促进HepG2细胞凋亡，抑制其增殖、迁移和侵袭能力，且较同浓度单药作用增强。

ObjectiveTo study the synergistic effects of curcumin and Cisplatin （DDP） on the proliferation, apoptosis, migration and invasion of hepatocellular carcinoma cells HepG2.MethodsHepG2 cells were randomly divided into multiple groups, including control [dmethyl sulfoxide （DMSO）], curcumin group （5, 10, 20 μmol/L）, DDP group （1, 5, 10 μmol/L） and combination treatment （Curcumin ＋ DDP） group （5 μmol/L Curcumin ＋ 5 μmol/L DDP, 10 μmol/L Curcumin＋ 5 μmol/L DDP）. The proliferation of HepG2 cells was detected by methyl thiazol tetrazolium （MTT） assay. Additionally, Hoechst 33258 assay was used to determine morphological changes of nucleus and cell apoptosis. The migration and invasion ability of HepG2 cells were detected by scratch healing assay and Transwell invasion assay, respectively.ResultsThe viability of HepG2 cells in combination treatment group was significantly lower than that in DDP group as indicated by MTT assay [（40.27±4.80）% vs. （61.80±9.14）%, P=0.023; （19.37±5.02）% vs. （61.80±9.14）%, P=0.002]. Hoechst 33258 staining showed that combination treatment triggered （10 μmol/L Curcumin＋ 5 μmol/L DDP） more severe cell apoptosis compared with DDP alone （5 μmol/L） [（18.95±1.30）% vs. （10.35±1.46）%, P=0.012]. Scratch healing assay revealed that curcumin or DDP alone induced significant inhibition of cell healing in a dose-dependent manner, while combination treatment exhibited more stronger effects on cell healing than curcumin alone [（33.81±1.63）% vs. （39.77±2.48）%, P=0.026; （24.54±4.93）% vs. （39.77±2.48）%, P=0.003]. Transwell invasion assay indicated that cell invasion was reduced under the treatment of curcumin or DDP, and combination treatment group significantly reduced the number of invasive cells more obvious （24.33±2.52 vs. 32.00±3.61, P=0.039; 15.67±3.06 vs. 32.00±3.61, P=0.004）.ConclusionCombination treatment could induce marked cell apoptosis of HepG2 and significantly inhibite cell proliferation, migration and invasion.