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目的观察榄香烯对人胃癌SGC-7901细胞株体外和体内增殖、抑制及凋亡的作用,并探讨其可能的作用机制。方法采用不同药物对培养后的人胃癌SGC-7901细胞株进行处理并分为4组:对照组、榄香烯组、细胞外信号调控的蛋白激酶1/2(ERK1/2)信号通路抑制剂PD98059组和榄香烯+PD98059联合组;再将20只雄性裸鼠随机分为4组:甲组(腹腔注射0.9%氯化钠注射液)、乙组(腹腔注射榄香烯最大耐受剂量200mg/kg)、丙组(腹腔注射PD98059最大耐受剂量1mg/kg)、丁组(腹腔注射最大耐受剂量榄香烯+PD98059),均制作胃癌移植瘤模型,并腹腔注射不同药物进行治疗。观察榄香烯和PD98059对细胞株相关蛋白的表达情况及裸鼠胃癌移植瘤的大小及对裸鼠生长的影响。采用MTT法检测细胞增殖情况,Western-blot法检测ERK1/2、磷酸化细胞外信号调控的蛋白激酶1/2(p-ERK1/2)、p38丝裂原活化蛋白激酶(p38)、磷酸化p38丝裂原活化蛋白激酶(p-p38)的表达,real-time RT-PCR检测Bcl-2 mRNA及Bax mRNA的表达,TUNEL法检测胃癌细胞凋亡并计算凋亡指数。结果与对照组相比,榄香烯组随药物浓度的增加,p-ERK1/2表达增加(P<0.05或0.01),Bax mRNA的表达增加(均P<0.01),Bcl-2 mRNA的表达降低[除榄香烯0.02mg/ml外,其他浓度均具有统计学差异(均P<0.01)],而总ERK1/2的表达无明显变化(均P>0.05);榄香烯、PD98059及榄香烯+PD98059对胃癌细胞增殖均具有抑制作用(均P<0.05),且两药联合的抑制作用最好;榄香烯、PD98059及榄香烯+PD98059组细胞凋亡率均高于对照组(P<0.05或0.01),并具有浓度依赖性,且两药联合组的凋亡率明显高于单一用药组(均P<0.05);乙、丁两组移植瘤的瘤重均较甲组明显减小(P<0.05或0.01),且两药联合时抑瘤率达75.59%。结论榄香烯对肿瘤细胞具有抑制作用,且可能是通过上调p-ERK1/2和p-p38蛋白的表达促进胃癌细胞和移植瘤的凋亡,另榄香烯与PD98059联合使用时对肿瘤细胞及组织具有协同抑制效应。
Objective To observe the effect of elemene on proliferation, apoptosis and apoptosis of human gastric cancer cell line SGC-7901 in vitro and in vivo and to explore its possible mechanism. Methods SGC-7901 cells were treated with different drugs and divided into four groups: control group, elemene group, inhibitor of extracellular signal-regulated protein kinase 1/2 (ERK1 / 2) signaling pathway PD98059 group and elemene + PD98059 combination group. Twenty male nude mice were randomly divided into 4 groups: group A (intraperitoneal injection of 0.9% sodium chloride injection), group B (intraperitoneal injection of the maximum tolerated dose of elemene 200mg / kg), group C (PD98059 maximal tolerated dose 1mg / kg) and group D (maximal tolerated dose of elemene + PD98059) were given gastric cancer xenograft model and intraperitoneal injection of different drugs for treatment . To observe the expression of elemene and PD98059 on cell lines and the size of nude mice bearing gastric cancer and their effects on the growth of nude mice. Cell proliferation was detected by MTT assay. The expressions of ERK1 / 2, p-ERK1 / 2, p38 mitogen-activated protein kinase (p38), phosphorylation p38 mitogen-activated protein kinase (p-p38), the expression of Bcl-2 mRNA and Bax mRNA were detected by real-time RT-PCR and the apoptosis index was calculated by TUNEL method. Results Compared with the control group, the expression of p-ERK1 / 2 increased (P <0.05 or 0.01), the expression of Bax mRNA increased (all P <0.01) and the expression of Bcl-2 mRNA (All P <0.01) except for elemene 0.02mg / ml, while the expression of total ERK1 / 2 had no significant change (all P> 0.05); Elemene, PD98059 and Elemene + PD98059 had inhibitory effects on the proliferation of gastric cancer cells (all P <0.05), and the combination of both drugs had the best inhibitory effect. The apoptosis rates of elemene, PD98059 and elemene + PD98059 group were higher than those of control (P <0.05 or 0.01). The apoptosis rates of the two groups were significantly higher than those of the single treatment group (all P <0.05) Group was significantly reduced (P <0.05 or 0.01), and the combined inhibition rate of two drugs reached 75.59%. Conclusion Elemene can inhibit tumor cells and may promote apoptosis of gastric cancer cells and xenografts by up-regulating the expressions of p-ERK1 / 2 and p-p38 proteins. When elemene is used in combination with PD98059, the tumor cells And the organization has a synergistic inhibitory effect.