加味生脉饮含药血清对非小细胞肺癌细胞增殖及迁移的影响

目的:研究加味生脉饮含药血清对非小细胞肺癌H460细胞增殖、细胞周期及其迁移的影响,并探讨其分子机制。方法:以生药量18 g·kg^-1 ig家兔,制备加味生脉饮含药血清,同容积生理盐水ig制备空白血清,并视血清浓度为100%。体外培养H460细胞,收集对数期生长的细胞,设置10%空白血清组,2.5%,5%,10%含药血清组,1 mg·L^-1 DDP组,共5组,调整细胞密度至3 000个/mL,血清作用24,48 h后,采用四甲基偶氮唑蓝(MTT)法测定细胞增殖抑制率;调整细胞密度至5×10⁵个/mL,加入上述浓度血清作用24 h后,分别采用流式细胞仪检测细胞周期,划痕实验观察细胞迁移的情况,Western blot法检测E钙连蛋白(E-cad)及波形蛋白(Vimentin)蛋白表达的变化。结果:与空白血清组相比,加味生脉饮含药血清作用H460细胞24 h后,细胞增殖抑制率上升(P<0.01),S期细胞百分率增高(P<0.01),划痕距离较长,E-cad蛋白含量升高(P<0.01),Vimentin蛋白含量降低(P<0.01)。结论:加味生脉饮对H460细胞增殖有一定抑制作用,主要途径为S期阻滞。加味生脉饮可以抑制H460细胞的转移,其机制可能与干预H460细胞上皮间质转化有关。

Effect of Drug-containing Serum with Modified Shengmai Decoction on Proliferation and Migration of Non Small Cell Lung Carcinoma Cells

Objective: To study the effect of drug-containing serum with modified Shengmai decoction on cell proliferation, cell cycle and migration of non small cell lung carcinoma(NSCLC)H460 cells, and investigate its molecular mechanisms. Method: Drug-containing serum with modified Shengmai decoction was prepared by administering the rabbits with 18 g·kg^-1 crude drug ig, and blank control serum was prepared with the same volume of normal saline(NS) ig. Both serum was regarded as a concentration of 100%. H460 cells were cultured in vitro and cells in logarithmic growth phase were collected. 10% control serum group, 2.5%, 5%, 10% drug-containing serum groups and 1 mg·L^-1 DDP group were established. Cell density was adjusted to 3 000 cells/mL. After serum action for 24, 48 hours, four methyl azo blue(MTT) method was used to measure the proliferation inhibition rate. Cell density was then adjusted to 5×10⁵ cells/mL, and after serum action for 24 hours, flow cytometry was used to test cell cycle, scratch assay was taken to observe cell migration, and Western blot method was used to measure the expression changes of E-cadherin(E-cad) and Vimentin. Result: Compared with the blank control group, drug-containing serum with modified Shengmai decoction had higher cell proliferation inhibition rate (P<0.01), higher cells percentage in S phase(P<0.01), longer wound scratch distance, higher protein content of E-cad and lower Vimentin (P<0.01) after serum action for 24 hours. Conclusion: Modified Shengmai decoction has an inhibition effect on cell H460 mainly by inhibiting cells in phase S. Modified Shengmai decoction can inhibit the metastasis of H460, whose mechanism may be associated with intervening the epithelial-mesenchymal transformation.