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【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 Yiqi Shutan Prescription on the proliferation of OCI-ly10 cells in diffuse large B-cell lymphoma and its mechanism. 【Methods】 CCK-8 (Cell Counting Kit-8) was used to detect the growth inhibition effect of Yiqi Shutan Prescription on OCI-ly10 cells. Flow cytometry was used to detect the changes of cell membrane phosphatidylserine fluorescein isothiocyanate / propidium iodide (Annexin V-FITC / PI) staining was used to detect the apoptosis of OCI-ly10 cells induced by Yiqi Chu Tan and Adriamycin. The effect of Yiqi Shutan Prescription on the expression of serine / threonine protein kinase (AKT) and phosphorylated AKT (Thr308, Ser473) in OCI-ly10 cells was detected by Western blot. [Results] The Yiqi Chu Tan Prescription-containing serum had inhibitory effect on OCI-ly10 cells in a certain time-concentration-dependent manner. Flow cytometry in the combination of Yiqiqitanfang containing serum and doxorubicin group of apoptosis rate, apoptosis rate of drug-containing doxorubicin group and drug-containing serum group than the negative control group was significantly (P <0.05). The effect of combination group on tumor cell apoptosis was significantly stronger than that of doxorubicin group and drug-containing serum group (all P <0.05). Western blot results showed that the Yiqi Chu Tan Decoction serum can significantly reduce the total AKT and p-AKT (Thr308) expression. 【Conclusion】 Yiqi Chu Tan Decoction can inhibit the proliferation and induce apoptosis of OCI-ly10 cells in diffuse large B-cell lymphoma, and has some antitumor and synergistic effects on doxorubicin, which may be related to its inhibitory effect AKT phosphorylation signaling pathway.