二氯乙酸钠和氯喹联合整合素亚单位的shRNA的协同抗肿瘤作用

目的:探究二氯乙酸钠、氯喹和表达整合素β3、β5亚单位的shRNA的重组病毒协同抗肿瘤作用。方法:探究二氯乙酸钠、氯喹和表达整合素β3、β5亚单位的shRNA的重组病毒对肝癌细胞株HepG2皮下异种移植瘤生长的影响,通过免疫组化试验检测药物对肿瘤血管生成、肿瘤细胞凋亡、增殖的影响。结果:在小鼠HepG2皮下移植瘤体内试验中可见单用氯喹对肿瘤生长抑制作用明显弱于单用二氯乙酸钠或两者合用,两药联合使用对肝癌细胞株HepG2产生的小鼠皮下移植瘤具有良好的抑制生长作用,并且使得小鼠能够很好地耐受氯喹,荷瘤所致的小鼠体重下降也得到缓解。单用表达整合素β3、β5亚单位的shRNA腺病毒对肿瘤抑制作用弱,合用两个化合物和表达整合素β3、β5亚单位的shRNA腺病毒则对小鼠HepG2移植瘤生长产生持续抑制作用,并减少单用表达整合素β3、β5亚单位的shRNA腺病毒对体重的影响。HE染色结果显示,给药后各组细胞结构被破坏,CD31和Ki67染色显示用药各组同步减少,TUNEL染色显示用药后各组细胞凋亡增加,其中二氯乙酸钠/氯喹/整合素β5和β3 shRNA腺病毒组最为明显,结果表明二氯乙酸钠/氯喹/整合素β5和β3 shRNA腺病毒联用具有显著的抗肿瘤生长作用,其机制是降低肿瘤的微血管密度、抑制肿瘤细胞的增殖和增加肿瘤细胞凋亡。结论:二氯乙酸钠、氯喹和表达整合素亚单位的shRNA的重组病毒进行组合具有协同抗肿瘤作用,为联合用药的设计及应用提供了参考。

Synergistic Anti-Tumor Effect of Sodium Dichloroacetate and Chloroquine and Recombinant Virus Expressing shRNA of Integrin Subunit

Objective:To explore the synergistic antitumor effect of sodium dichloroacetate and chloroquine and recombinant adenovirus expressing shRNA of integrin subunitβ3,β5.Methods:Subcutaneous xenografts of hepatocellular carcinoma cell line HepG2 was established to detect the inhibitory effect of recombinant virus expressing shRNA of integrin subunitβ3,β5 combined with sodium dichloroacetate and chloroquine in vivo.Immuno-histochemical experiments were further performed to investigate the effects of the combination on tumor angiogenesis,tumor cell apoptosis and proliferation.Results:Chloroquine alone demonstrated weaker inhibitory effects on growth of transplanted tumor of hepatocellular carcinoma cell line HepG2 than sodium dichloroacetate or the combination of sodium dichloroacetate and chloroquine.And the mice can tolerate chloroquine well.Moreover,the loss of weight caused by the tumor-bearing mice was also alleviated.Adenovirus expressing shRNA of integrin subunitβ3,β5 also showed little inhibitory effect on tumor growth.However,the combination of sodium dichloroacetate and chloroquine with the adenovirus expressing integrin shRNA significantly inhibited the tumor growth.Moreover,the effect of adenovirus expressing integrin shRNA on mouse weight was attenuated by the combination.The results of HE staining showed that the cell structure of each group has been damaged after administration.CD31 and Ki67 staining showed that the proliferation of tumor cells was reduced simultaneously after administration.TUNEL staining showed that apoptosis of each group increased after administration.Among them,the combination of sodium dichloroacetate,chloroquine and recombinant virus expressing shRNA of integrin subunitβ3,β5 was the most significant.The results demonstrated that the combination of sodium dichloroacetate/chloroquine/integrinβ3 andβ5 shRNA adenovirus had a significant anti-tumor effect.And its mechanism is to reduce tumor micro-vessel density,inhibit tumor cell proliferation and increase tumor cell apoptosis.Conclusions:Sodium dichloroacetate and chloroquine combined with adenovirus expressing shRNA of integrin subunitβ3,β5 can obtain a better anti-tumor effect.The study provides new insights into the design and application of the combined medicine.