赛来昔布联合吉西他滨对胰腺癌细胞侵袭转移的抑制作用及其机制

Inhibition Of invasive potential of pancreatic carcinoma cells by celecoxib in combination with gemcitabine

目的研究环氧合酶-2选择性抑制剂赛来昔布联合吉西他滨对胰腺癌细胞侵袭力的抑制作用及其机制。方法采用体外侵袭力测定试剂盒和鸡胚尿囊膜模型检测赛来昔布联合吉西他滨对胰腺癌细胞侵袭力的影响，应用RT-PCR方法检测赛来昔布和吉西他滨对胰腺癌细胞基质金属蛋白酶MMP-2、MMP-9及其组织抑制剂TIMP-1、TIMP-2mRNA表达情况，应用明胶酶谱和反式酶谱方法检测赛来昔布和吉西他滨对胰腺癌细胞分泌MMP-2、MMP-9、TIMP-1及TIMP-2的影响。结果吉西他滨作用24h和72h胰腺癌侵袭细胞数量较对照组无明显差异，赛来昔布作用24h即可降低穿透ECM膜的侵袭细胞数量，赛来昔布与吉西他滨联合，侵袭细胞数较对照组明显减少，但较单用赛来昔布无明显差异，随着作用时间的延长，侵袭细胞数有下降趋势。药物作用24h时，对鸡胚胰腺癌移植瘤细胞的侵袭无明显抑制作用，吉西他滨作用72h，鸡胚胰腺癌移植瘤细胞侵袭未受抑制，赛来昔布作用72h，鸡胚胰腺癌移植瘤细胞侵袭明显受抑制，两种药物联合作用72h，对鸡胚移植瘤细胞侵袭力的抑制作用与单用赛来昔布无明显差异。RT-PCR、明胶酶谱和反式明胶酶谱分析表明，赛来昔布抑制胰腺癌细胞分泌和激活MMP-2和MMP-9，但对TIMP-1和TIMP-2的分泌和激活无明显影响，联合吉西他滨前后，MMP-2、MMP-9、TIMP-1和TIMP-2的分泌和活性无明显改变。结论COX-2选择性抑制剂赛来昔布抑制吉西他滨干预后的胰腺癌的细胞侵袭，从而间接对吉西他滨的治疗起增敏作用，其机制部分通过是抑制MMP-2和MMP-9的分泌，减少ECM的降解所致。

Objective To investigate the inhibitory effect of celecoxib, a selective cyclooxygenase-2 inhibitor, in combination with gemcitabine on invasive potential of pancreatic carcinoma cells. Methods In vitro cell invasion assay and chorioallantoic membrane （CAM） grafted model were used to evaluate the invasive potential of the pancreatic carcinoma cells. The expression of MMP-2, MMP- 9, TIMP-1 and TIMP-2 was determined by RT-PCR. Conditional medium was examined for MMP-2, MMP-9, TIMP-1 and TIMP-2 by zymography and reverse zymography. Results In vitro and in vivo invasion of SW1990 cells was inhibited by celecoxib but had no significant change in gemcitabine as compared with the control group. Celecoxib in combination with gemcitabine compressed the invasive potential of SW1990 cells but had no marked difference from celecoxib. The expression and secretion of MMP-2 and MMP-9 was closely related to the changes in cell invasive potential induced by different drugs. However, the expression of TIMP-1 and TIMP-2 did not significantly alter in all the 4 groups. Conclusions Celecoxib can effectively inhibit the invasion of pancreatic carcinoma cells pretreated by gexneitabine. The sensitization effect of celecoxib on gemcitabine is indirect and inhibition of the MMP- 2 and MMP-9 secretion is involved in this process.