仙连解毒方对结直肠癌原位瘤小鼠肿瘤增殖的抑制作用及机制探讨

Inhibitory effect and mechanism investigation of Xianlian Jiedu Recipe on tumor proliferation in the model of colorectal cancer in situ mice

目的:探讨仙连解毒方对结直肠癌(CRC)原位瘤小鼠肿瘤的抑瘤作用,并对其作用机制进行探讨。方法:建立CT26原位瘤小鼠模型,按体质量随机分为正常组、模型组、5-FU给药组、仙连解毒方给药组,连续给药28 d,隔日观察小鼠状态。处死小鼠后剥取瘤组织,HE染色观察组织病理形态学变化;免疫组织化学染色法检测小鼠瘤组织增殖细胞核抗原(PCNA)表达;TUNEL染色观察仙连解毒方对原位肿瘤细胞凋亡的影响;Western Blot检测原位瘤组织内糖原合成酶激酶-3β(GSK-3β)、p-GSK-3β、β-连环蛋白(β-catenin)蛋白表达的变化。结果:与模型组比较,仙连解毒方可以显著减少原位瘤的体积(P<0.01),HE及TUNEL染色结果显示仙连解毒方使肿瘤组织凋亡及坏死细胞增加;免疫组织化学染色结果显示,仙连解毒方给药组小鼠瘤组织内PCNA的蛋白表达减少。Western Blot染色结果显示,仙连解毒方给药后小鼠瘤组织内p-GSK-3β/GSK-3β比值显著下降(P<0.01),β-catenin蛋白表达显著下调(P<0.01)。结论:仙连解毒方具有抑制小鼠CRC原位瘤增殖的作用,其作用机制与干预Wnt/β-catenin通路,抑制肿瘤细胞的增殖、促进肿瘤细胞凋亡有关。

Objective:To explore the anti-tumor effect and the possible mechanism of Xianlian Jiedu Recipe on colorectal cancer(CRC)in situ mice.Methods:After established the CT26 cells tumor-bearing model,all the mouse were divided into four groups:normal group,model group,5-FU group,Xianlian Jiedu Recipe group,continuous administration was lasted for 28 days,and the growth of tumor were observed every other day.After administration,the tumor tissues were sacrificed and the histopathological changes were observed by HE staining.The expression of PCNA in tumor tissues was detected by immunohistochemical staining.TUNEL staining was used to observe the effect of Xianlian Jiedu Recipe on apoptosis of in situ tumor cells.The expression of GSK-3β,p-GSK-3β,β-catenin in the tumor tissues was detected by Western Blot.Results:Compared with the model group,Xianlian Jiedu Recipe group could obviously decrease the volume of tumor(P<0.01).HE and immunohistochemical staining showed that the necrosis cells increased in tumor tissue and the expression of PCNA was decreased in Xiaolian Jiedu Recipe group.Western Blot staining showed that after administration of Xianlian Jiedu Recipe,the ratio of p-GSK-3β/GSK-3βwas decreased significantly(P<0.01),the protein expression ofβ-catenin was also down regulated(P<0.01).Conclusion:Xianlian Jiedu Recipe can inhibite the proliferation of tumor tissue in situ mice,and its mechanism is related to the intervention of Wnt/β-catenin pathway,inhibition of tumor cell proliferation and promotion of tumor cell apoptosis.