原卟啉钠体外对人肝癌细胞株SMMC-7721细胞的抑制作用

目的:研究原卟啉钠(NAPP)体外对人肝癌细胞株SMMC-7721细胞的抑制作用。方法:通过对人正常肝细胞株QSG-7701细胞的药物毒性实验筛选出NAPP对人正常肝细胞无明显毒性的最大无毒浓度(TC0)。以TC0为起始浓度,10倍梯度稀释成5个不同的NAPP浓度,作用于体外培养的人肝癌细胞株SMMC-7721细胞,通过MTT比色法测定NAPP对SMMC-7721细胞的增殖抑制作用;经倒置光学显微镜和Hoechst33258染色倒置荧光显微镜进行形态学观察,计数凋亡细胞,并计算凋亡指数(AI);采用AnnexinⅤ-FITC/PI双标流式凋亡检测试剂盒经流式细胞仪检测SMMC-7721细胞凋亡和坏死率。结果:不同浓度NAPP作用后,SMMC-7721细胞的增殖明显受到抑制,A值明显降低(P<0.05或P<0.01),且抑制率呈一定的浓度-时间依赖性;光学显微镜下见细胞个数明显减少,细胞密度明显变稀,细胞形态发生了明显变化,很多细胞间的连接消失,细胞变圆、皱缩或见细胞外形变长呈长梭形;荧光显微镜下细胞呈典型的凋亡形态学改变,AI显著升高(P<0.05或P<0.01);经流式细胞仪检测,可见SMMC-7721细胞凋亡和坏死率不同程度升高。结论:NAPP在体外对人肝癌细胞株SMMC-7721细胞具有一定的抑制增殖、促进凋亡和坏死的作用。

Inhibitory Effect of Protoporphyrin Disodium on Human Hepatoma Carcinoma Cell Strain of SMMC-7721 Cells in Vitro

Objective: To study the inhibitory effect of protoporphyrin disodium (NAPP) on human hepatoma carcinoma cell strain of SMMC-7721 cells in vitro . Method: The maximum atoxic concentration (TC₀) of NAPP was determined by drug toxicity test on normal human hepatocyte strain of QSG-7701 cells. TC₀ was used as the initial concentration, five different concentrations of NAPP were diluted by 10 times gradient dilution, and applied to treat the human hepatoma carcinoma cell strain of SMMC-7721 cells cultured in vitro . MTT assay was used for evaluating the proliferation inhibitory effect of NAPP. Morphological observation for SMMC-7721 cells was undertaken via the inverted optical microscope and inverted fluorescence microscope after Hoechst33258 staining. Apoptotic cells were counted and apoptosis index (AI) was calculated. Flow cytometry was applied to measure the apoptosis and necrosis rates of SMMC-7721 cells through Annexin V-FITC/PI double staining with apoptosis detection kit. Result: The proliferation of SMMC-7721 cells was inhibited dramatically after the treatment of different concentrations of NAPP, and the A values were decreased significantly (P<0.05 orP<0.01). The inhibition rate was depended on the concentration of NAPP and the treatment time to some extent. These were visible in the optical microscope that cell number reduced obviously, cell density was thinning markedly, the appearance of SMMC-7721 cells was changed evidently, significantly, many conjunctions between cells disappeared, and cells became round and corrugativus or cellular shape turned slender like Fusiform shape. The SMMC-7721 cell nucleus showed the typical changes of apoptosis morphology in fluorescence microscope, and AI increased remarkably (P<0.05 orP<0.01). Flow cytometry showed that apoptosis and necrosis rates of SMMC-7721 cells were raised to different level. Conclusion: NAPP can inhibit the proliferation of human hepatoma carcinoma cell strain of SMMC-7721 cells and promote its apoptosis and necrosis.