益气除痰方对弥漫大B细胞淋巴瘤增殖及AKT表达的影响

Effect of Qi-benefiting and Phlegm-eliminating Recipe on Proliferation and Serine/threonine-protein Kinase AKT Expression of Diffused Large B-cell Lymphoma Cells

【目的】研究益气除痰方对弥漫大B细胞淋巴瘤OCI-ly10细胞增殖的影响及其作用机制。【方法】应用CCK-8(Cell Counting Kit-8)法检测益气除痰方对OCI-ly10细胞的生长抑制作用,以流式细胞术细胞膜磷脂酰丝氨酸异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)染色检测联合应用益气除痰方和阿霉素诱导OCI-ly10细胞凋亡的能力。以Western blot法检测益气除痰方对OCI-ly10细胞丝氨酸/苏氨酸蛋白激酶(AKT)及磷酸化AKT(Thr308、Ser473)表达的影响。【结果】益气除痰方含药血清对OCI-ly10细胞具有生长抑制作用,且具有一定的时间—浓度依赖性。流式细胞术检测中,联合应用益气除痰方含药血清和阿霉素组的凋亡率、阿霉素单药组凋亡率及含药血清组凋亡率均较阴性对照组显著升高,其中联合组促进肿瘤细胞凋亡的作用显著强于阿霉素单药组及含药血清组(均P<0.05)。Western blot结果显示益气除痰方含药血清可显著降低总AKT和p-AKT(Thr308)的表达。【结论】益气除痰方对弥漫大B细胞淋巴瘤OCI-ly10细胞具有生长抑制及促进凋亡的作用,并对阿霉素具有一定的抗肿瘤增效作用,其作用机制可能与抑制了AKT磷酸化的信号通路相关。

Objective To investigate the effect of Qi-benefiting and Phlegm-eliminating Recipe （QPR） on the proliferation of diffused large B-cell lymphoma cells （OCI-lyl0） and to explore its mechanism. Methods The inhibitory effect of QPR on the proliferation of OCI-lyl0 cells was detected by Cell Counting Kit-8（CCK8）. The apoptosis of OCI-lyl0 induced by QRP and doxorubicin was determined by flow cytometry combined with Annexin V-fluorescein isothiocyanate/propidium iodide （FITC/PI） staining method. The effect of QPR on the expression of serine/threonine protein kinase AKT and phosphorylated-AKT （Thr308 and Ser 473）was evaluated by Western blotting method. Results The growth of OCI-lyl0 was inhibited by QPR-containing serum in time- concentration-dependent manner. The apoptotic rate o[ combination group（treated by QPR-containing serum and doxorubicin） , doxorubicin group and QPR-containing serum group was significantly higher than that of the negative control group, and the combination group had the highest apoptotic rate （P〈0.05）. The results of Western blotting showed that AKT and p-AKT（Thr308） expression levels were down-regulated by QPR- containing serum（P〈0.05 ）. Conclusion QPR is effective on inhibiting OCI-lyl0 growth and promoting OCI-lyl0 apoptosis, and can enhance the anti-tumor effect of doxorubicin. Its possible mechanism may be associated with the inhibition of p-AKT related signaling pathway.