青龙衣含药血清抑制胃癌SGC-7901细胞生长和诱导凋亡

Serum Containing Juglans mandshurica Pericarp Inhibits Proliferation and Induces Apoptosis of Gastric Carcinoma SGC-7901 Cell

摘要:

目的：探讨青龙衣含药血清对胃癌SGC-7901细胞增殖及凋亡的影响，并探讨其作用机制。方法：Wistar大鼠随机分为空白组、青龙衣低剂量（25 g·kg^-1·d^-1），中剂量（50 g·kg^-1·d^-1），高剂量（100 g·kg^-1·d^-1）组及顺铂（5 g·kg·d^-1）组，连续灌胃给药7 d，空白组灌胃给予等体积生理盐水；末次给药2 h后，颈动脉采血，制备青龙衣含药血清；分别采用噻唑蓝（MTT）比色法、流式细胞术、烟酸己可碱（Hoechst）染色、实时荧光定量聚合酶链式反应（Real-time PCR）及蛋白免疫印迹法（Western blot）检测青龙衣含药血清对SGC-7901细胞增殖、凋亡率、凋亡指数、凋亡相关基因及凋亡信号通路蛋白表达的影响。结果：与空白组比较，青龙衣含药血清对SGC-7901细胞的增殖具有明显抑制作用（P<0.05），青龙衣含药血清处理48 h后的早期、晚期凋亡率，凋亡指数及促凋亡基因Bcl-2相关X蛋白（Bax），Bcl-2相关K蛋白（Bak）mRNA表达均升高，抗凋亡基因B淋巴细胞瘤-2（Bcl-2）mRNA表达降低（P<0.05）；青龙衣含药血清组β-链蛋白（β-catenin），细胞周期蛋白D₁（Cyclin D₁）及原癌基因（C-myc）蛋白表达均明显降低（P<0.05）。结论：青龙衣含药血清对胃癌SGC-7901细胞的增殖具有抑制作用，该作用可能与诱导凋亡及抑制分泌型糖蛋白（Wnt）/β-catenin通路的活化有关。

Abstract:

Objective: To explore the effects of serum containing Juglans mandshurica (SCJM) pericarp on proliferation and apoptosis of gastric carcinoma SGC-7901 cells. Method: Wistar rats were randomly divided into blank control group, low (25 g·kg^-1·d^-1), medium (50 g·kg^-1·d^-1) and high (100 g·kg^-1·d^-1) dose J.mandshurica treatment groups and cisplatin group (5 g·kg^-1·d^-1). The rats in treatment groups received lavage administration for 7 d, and the rats in blank control group received the equivalent volume of normal saline. 2 h after the last treatment, SCJM was prepared by carotid blood serum; cell proliferation, apoptosis rates, apoptosis index, levels of apoptosis related gene and protein expression were evaluated by methylthiazolyldiphenyl-tetrazolium bromide (MTT) colorimetry, flow cytometry, Hoechst staining, Real-time PCR and Western blot respectively. Result: SCJM could inhibit the proliferation of SGC-7901 cells (P<0.05) as compared with blank group, and 48 h after treatment by SCJM, early-and late-apoptosis rates, apoptosis index and mRNA levels of Bax and Bak were increased, but mRNA level of B-cell lymphoma-2 (Bcl-2) was decreased (P<0.05); the protein levels of β-catenin, Cyclin D₁ and C-myc in SCJM-treatment groups were lower than those of control group (P<0.05). Conclusion: SCJM had inhibitory effect on proliferation of gastric carcinoma SGC-7901 cells, and the effect may be related to the induction of apoptosis and inhibition of Wnt/β-catenin pathway activation.