金龙蛇颗粒对裸鼠原位移植人胃癌MKN-45细胞凋亡的影响

Effects of Jinlongshe Granules on apoptosis of MKN-45 human gastric cancer cells orthotopically transplanted in nude mice

目的：评价金龙蛇颗粒对裸鼠原位移植入胃癌MKN-45细胞的抑制作用.方法：50只裸鼠建立原位移植人胃癌MKN-45细胞模型,随机分为模型组,金龙蛇颗粒高、中、低剂量组和5-FU干预组.各组裸鼠予以相应药物实施干预.观察各组荷瘤裸鼠一般情况,肿瘤生长情况及抑瘤率.流式细胞仪检测肿瘤细胞周期分布及细胞凋亡情况.采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-fluorescein isothiocyanate/propidium iodide,Annexin V-FITC/PI）双标记染色法鉴别早期凋亡细胞、晚期凋亡细胞和坏死细胞.电镜下观察肿瘤细胞的超微结构变化.结果：金龙蛇颗粒高、中、低剂量组的抑瘤率分别为68.13%、55.94%和50.31%,呈剂量依赖效应,与5-FU干预组抑瘤率比较,差异无统计学意义.金龙蛇颗粒各剂量组肿瘤细胞凋亡率为22.81%～38.54%,亦呈剂量依赖效应,且肿瘤细胞主要被阻滞于G0/G1期.AnnexinV-FITC/PI双标记染色结果显示,金龙蛇颗粒各剂量组以早期凋亡细胞居多,5-FU干预组则以晚期凋亡细胞和坏死细胞居多.结论：金龙蛇颗粒对裸鼠原位移植人胃癌MKN-45细胞具有较好的抑制作用,促进MKN-45细胞凋亡可能是其抗肿瘤治疗的主要作用机制之一.

Objective： To assess the effects of Jinlongshe Granules （JLSG） on tumor growth of gastric carcinoma. Methods： Fifty nude mice orthotopically transplanted with MKN-45 human gastric cancer cells were divided into five groups： untreated group, 5-fluorouracil （5-FU）-treated group and high-, medium-, and low-dose JLSG-treated groups. Corresponding antitumor drugs were administered in each group except the untreated group. The antitumor effects in vivo were evaluated. Cell cycle distribution and apoptosis of MKN- 45 human gastric cancer cells were determined by using flow cytometry （FCM） and Annexin V-fluorescein isothiocyanate/propidium iodide （Annexin V-FITC/PI） staining assay. The ultrastructure of MKN-45 gastric cancer cells was observed by transmission electron microscope. Results： In the mice treated with high-, medium-, and low-dose JLSG, the growth inhibition rates of gastric cancer were 68.13 %, 55.94 % and 50.31% respectively,＇ and this antitumor effect was dose-dependent. In the mice treated with intraperitoneal injection of 5-FU, the growth inhibition rate of gastric cancer was 53.43% and not much different from those treated with JLSG. The apoptotic rates in the high-, medium-, and low-dose JLSG-treated groups were 22.81%, 28.27% and 38. 54% respectively, in a dose-dependent manner, with the cell cycle arrested at G0/G1 phase. An Annexin V-FITC/PI staining assay revealed that the percentages of early apoptotic cells in the three dose JLSG-treated groups were all significantly higher than that in the 5-FU-treated group, whereas the late apoptotic and necrotic cells were much more in the 5-FU-treated group than those in the three dose JLSGtreated groups. Conclusion： Jinlongshe Granules exert an inhibiting effect on MKN-45 human gastric cancer cells orthotopically transplanted in nude mice by promoting the cell apoptosis.