益气扶正复方联合依维莫司对三阴性乳腺癌MDA-MB-468细胞株裸鼠移植瘤的影响

目的研究益气扶正复方联合依维莫司对三阴性乳腺癌MDA-MB-468细胞株裸鼠移植瘤的抑制作用及其作用机制。方法体外培养三阴性乳腺癌MDA-MB-468细胞株,将细胞悬液接种于雌性BALB/C裸鼠第二乳腺乳垫处,建立皮下移植瘤模型。将造模成功的裸鼠随机分为对照组、益气扶正复方(中药复方)组(40mg/kg)、依维莫司组(2mg/kg)和联合组,每组6只。各组灌胃给予相应药物干预,连续30d。比较各组移植瘤体积和质量的变化,计算各组肿瘤抑制率。采用Real-time PCR检测各组mTor、Akt、P70s6k、4Ebp-1 mRNA的表达水平,Western blot检测各组Akt、p-Akt、m-TOR、p-mTOR、PTEN及mTOR下游底物p-4EBP-1、p-P70S6K的蛋白表达水平。结果与中药复方组相比,联合组移植瘤体积和质量显著减小(P0.05,P0.01),肿瘤抑制率明显升高,且联合组mTor和P70s6k mRNA表达及p-mTOR和P70S6K蛋白表达水平明显低于中药复方组,而联合组PTEN蛋白表达高于中药复方组(P0.05)。结论益气扶正复方与依莫维司联用能抑制MDA-MB-468荷瘤裸鼠的肿瘤组织生长,与益气扶正复方单用相比效应更为显著,其作用机制可能与抑制p-mTOR、p-Akt、p-P70S6K活性,提高p-4EBP-1、PTEN表达有关。

Effects of Yiqi Fuzheng Compound Recipe combined with everolimus on xenograft of triple negative breast cancer MDA-MB-468 cell line in nude mice

  Objective: To study the inhibitory effects and mechanism of Yiqi Fuzheng Compound Recipe combined with everolimus on xenograft of nude mice with triple negative breast cancer MDA-MB-468 cell line. MethodsThe triple negative breast cancer cell line MDA-MB-468 was cultured in vitro and the cell suspension was inoculated on the second breast pad of female BALB/C nude mice to establish a subcutaneous xenograft model. The successful modeling nude mice were randomly divided into the control group, Yiqi Fuzheng Compound Recipe (Chinese medicine compound recipe) group(40 mg/kg), everolimus group(2 mg/kg)and combination group, 6 mice in each group. Each group was treated with the corresponding drugs by intragastric administration for continuous 30 days. The volume and weight of transplanted tumor in each group were compared, and the tumor inhibition rate was calculated. The mRNA expressions of mTor, Akt, P70s6k and 4Ebp-1 in each group was detected by Real-time PCR. The protein expressions of Akt, p-Akt, m-TOR, p-mTOR, PTEN and p-4EBP-1 and p-P70S6K(as mTOR downstream substrates)were detected by Western blot.  Results:Compared with the Chinese medicine compound recipe group, the volume and weight of transplanted tumor in the combination group were decreased significantly(P<0.05,P<0.01), and the tumor inhibition rate was obviously increased. The mRNA expressions of mTor and P70s6k and the protein expressions of p-mTOR and P70S6K in the combination group were obviously lower than those in the Chinese medicine compound recipe group, and the protein expression of PTEN in the combination group was higher than that in the Chinese medicine compound recipe group(P<0.05).  Conclusion:The combination of Yiqi Fuzheng Compound Recipe and everolimus can inhibit the growth of tumor tissue in nude mice bearing MDA-MB-468 cell, and the effects are more significant compared with Yiqi Fuzheng Compound Recipe alone. The mechanism may be related to inhibiting the activities of p-mTOR, p-Akt, p-P70S6K and increasing the expressions of p-4EBP-1 and PTEN.