摘要
目的体外研究选择性环氧合酶-2抑制剂塞来昔布联合热疗对人胰腺癌SW1990细胞的协同抑制作用。方法四甲基偶氮唑盐（MTT）法研究热疗、塞来昔布及联合应用时对细胞增殖的抑制作用。电镜和流式细胞仪下检测不同处理因素对细胞凋亡和细胞周期的影响。逆转录聚合酶链式反应（RT—PCR）法检测bax、p16、p27^Kipl、热休克蛋白（HSP）70的表达。Western印迹法检测HSP70蛋白的表达。结果MTT法示热疗组、塞来昔布组、联合组的细胞存活率分别为56．63％、44．55％、21．83％。电镜、流式细胞仪检测结果示三种作用均可诱导细胞凋亡，联合组、塞来昔布组、热疗组及对照组凋亡比例分别为33．32％、17．33％、11．78％、6．66％，联合组对细胞周期的阻滞作用最明显。RT—PCR示各处理组中p27^Kipl、bax表达明显上调，尤以联合组最为明显；p16、HSP70在热疗组和联合组中表达明显上调；Western印迹法示HSP70在各组细胞中都有表达，但在热疗组和联合组中表达更强。结论塞来昔布联合热疗对胰腺癌SW1990细胞生长具有协同抑制效应。两者联用可能通过上调HSP70、bax、p16和p27^Kipl的表达诱导细胞凋亡和细胞周期停滞。
Objective To study the synergistic effect of celebrex, a kind of selective cyclooxygenase-2 inhibitor, combined with thermotherapy on the growth of human pancreatic carcinoma SW1990 cells in vitro, and its possible mechanism. Methods SW1990 cells were treated with thermotherapy or celebrex or in their combination. Anti-proliferative effect was evaluated by MTT assay. Electron microscopy and flow cytometry were used to observe the apoptosis and the cell cycles. Heat shock protein（HSP）70, p16, bax, p27^Kipl were measured by RT-PCR. The expression of HSP70 was measured by Western blot. Results MTT assay and flow cytometry demonstrated that combination group had the strongest inhibitory effect on the growth of SW1990 cells as well as cell cycle G1 phase stagnancy and S phase proportion reduction. Apoptosis could be observed in all treated groups through electron microscopy. But the apoptosis ratios was different in these groups： combination group〉 celebrex group〉thermotherapy group〉 control group. RT-PCR showed p27^Kipl and bax was up regulated in all managed groups, especially in combination group. The expression of p16 and HSP70 significantly increased in thermotherapy and combination groups. Similar trend of HSP70 expression was showed by Western blot. Conclusions Celebrex combined with thermotherapy have a synergistic inhibitor effect on the growth of human pancreatic carcinoma in vitro. Thermotherapy combined with celebrex may increase the expression of HSP70, bax, p16 and p27^Kipl so as to induce tumor cell apoptosis as well as to induce G1 phase stagnancy.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2007年第1期23-26,共4页
Chinese Journal of Digestion
