汉防己甲素对口腔癌小鼠血清TGF-β1和PDGF水平的影响及相关机制研究

Effects of tetrandrine on serum TGF-β1 and PDGF levels in mice with oral cancer and related mechanisms

目的探究汉防己甲素对口腔癌小鼠血清转化生长因子-β1(TGF-β1)和血小板衍生生长因子(PDGF)水平的影响及相关机制。方法以30只雄性C57BL/6小鼠为研究对象,通过在小鼠腋窝皮下注射20μL的人口腔鳞状细胞癌Tca8113细胞悬液建立小鼠人口腔癌异种移植瘤模型,将小鼠根据实验目的分为模型组和汉防己甲素治疗组,各15只。通过小鼠酶联免疫吸附试验试剂盒检测其血清TGF-β1和PDGF水平。通过聚合酶链式反应检测小鼠移植瘤组织TGF-β1和PDGF的mRNA表达。通过免疫组织化学染色评估肿瘤组织的血管密度和血管周细胞覆盖率。通过TUNEL和免疫染色分析肿瘤细胞的凋亡、活力。测量肿瘤的最大直径和最大横向直径并计算体积。通过蛋白印迹分析小鼠肿瘤组织Erk1/2、Akt、Smad 2和Smad 3的蛋白表达。结果治疗组较模型组血清TGF-β1和PDGF水平降低,差异均有统计学意义(P<0.05)。治疗组较模型组肿瘤组织中TGF-β1和PDGF mRNA表达降低,差异均有统计学意义(P<0.05)。治疗组较模型组的血管密度和血管周细胞覆盖率降低,差异均有统计学意义(P<0.05)。治疗组较模型组细胞凋亡升高,细胞活力降低,差异均有统计学意义(P<0.05)。第1周两组小鼠肿瘤体积比较差异无统计学意义(P>0.05),第2周和第4周,治疗组较模型组体积减小,差异均有统计学意义(P<0.05)。治疗组较模型组Erk1/2、Akt、Smad 2和Smad 3的蛋白表达降低,差异均有统计学意义(P<0.05)。结论汉防己甲素通过PDGF/TGF-β1/Smad2/3信号通路下调口腔癌小鼠血清TGF-β1和PDGF水平,抑制异种移植肿瘤新血管形成,促进肿瘤细胞的凋亡,从而起到抗癌作用。

Objective To explore the effect of tetrandrine on serum transforming growth factor-β1(TGF-β1)and platelet-derived growth factor(PDGF)levels in oral cancer mice and related mechanisms.Methods Thirty male C57BL/6 mice were used as research objects.A mouse human oral cancer xenograft model was established by subcutaneously injecting 20μL of human oral squamous cell carcinoma Tca8113 cell suspension into the mouse armpit.The mice were divided into model group and tetrandrine treatment group according to the experimental purpose,each with 15 mice.The serum TGF-β1 and PDGF levels were detected by mouse enzyme-linked immunosorbent assay kit.The mRNA expression of TGF-β1 and PDGF in mouse transplanted tumor tissue was detected by PCR.The vascular density and perivascular cell coverage of tumor tissues were evaluated by immunohistochemical staining.The apoptosis and viability of tumor cells were analyzed by TUNEL and immunostaining.Measure the maximum diameter and maximum lateral diameter of the tumor and calculate the volume.The protein expression of Erk1/2,Akt,Smad 2 and Smad 3 in mouse tumor tissues was analyzed by Western blotting.Results The serum TGF-β1 and PDGF levels in the treatment group were lower than those in the model group,the differences were statistically significant(P<0.05).The expression of TGF-β1 and PDGF mRNA in the treatment group was lower than that in the model group,the differences were statistically significant(P<0.05).Compared with the model group,the treatment group had lower blood vessel density and pericyte coverage,the differences were statistically significant(P<0.05).Compared with the model group,the treatment group had higher cell apoptosis and lower cell viability,the differences were statistically significant(P<0.05).There was no difference in tumor volume between the two groups in the first week(P>0.05).In the second and fourth weeks,the volume of the treatment group was smaller than that of the model group,the differences were statistically significant(P<0.05).The protein expression of Erk1/2,Akt,Smad 2 and Smad 3 in the treatment group was lower than that in the model group,the differences were statistically significant(P<0.05).Conclusion Tetrandrine down-regulates serum TGF-β1 and PDGF levels in oral cancer mice through PDGF/TGF-β1/Smad2/3 signaling,inhibits the formation of new blood vessels in xenograft tumors,promotes tumor cell apoptosis,and plays an anti-cancer effect.