干扰EMMPRIN表达对胰腺星状细胞增殖及侵袭能力的影响  

Silencing EMMPRIN inhibits the proliferation and infiltrations of pancreatic stellate cells

作　　者：张巍巍 李林浩[2] 战淑慧 张东生[3] 耿长新 孙昕 谢方瑜[4] 宋敏[4] 赵丽萍 解祥军 

机构地区：[1]青岛市立医院消化科,山东省266071 [2]青岛市立医院普外科 [3]青岛市立医院肝胆外科 [4]青岛市立医院心内科

出　　处：《中华普通外科杂志》2015年第11期885-888,共4页Chinese Journal of General Surgery

摘　　要：目的探讨RNA干扰细胞外基质金属蛋白酶诱导分子（extracellular matrix metalloproteinase inducer，EMMPRIN）的表达对胰腺星状细胞（pancreatic stellate cell，PSC）增殖、侵袭及转移能力的影响。方法应用QRT-PCR法及细胞免疫组化法检测EMMPRIN在PSC中mRNA含量及蛋白的表达；PSC经RNA干扰后，Westernblot法检测EMMPRIN在PSC中封闭效果，MTr、软琼脂法检测细胞增殖能力，Transwell法检测EMMPRIN封闭前后PSC（PSC-NC，PSC-EMMPRINi）细胞侵袭能力，ELISA法检测MMPRIN封闭前后PSC（PSC-NC，PSC-EMMPRINi）及与ASPC细胞联合培养后（ASPC-NC／PSC-NC，ASPC-EMMPRINi／PSC-EMMPRINi）培养基中MMP2、VEGF的分泌量。结果PSC中EMMPRINmRNA呈高表达（1848．67±281．01）copies／μl，EMMPRIN蛋白主要定位于PSC的细胞膜及细胞质中，PSC经RNA干扰后，EMMPRIN蛋白在PSC中下调70％～90％，EMMPRIN封闭前后PSC增殖能力差异无统计学意义（P=0．199），EMMPRIN封闭后PSC-EMMPRINi及联合培养ASPC-EMMPRINi／PSC．EMMPRINi的培养基中MMP2分泌量较封闭前PSC-NC及ASPC-NC／PSC-NC稍减少（P：0．16），PSC-EMMPRINi较PSC-NC侵袭能力显著降低（P=0．046）。结论干扰EMMPRIN在PSC中的表达可抑制其侵袭侵润能力。Objective To investigate the role of extracellular matrix metalloproteinase inducer molecule （EMMPRIN） on proliferative, invasive and metastatic ability of pancreatic stellate ceils （PSC） by silencing EMMPRIN with RNA interferance. Methods QRT-PCR and immunocytochemistry were applied to detect the expression of EMMPRIN mRNA and protein in PSC; Western blot assay was used to test EMMPRIN expression of PSC blocked by RNA interference; MTT and soft agar assay were used to test PSC proliferative ability, and transwell assay to measure PSC invasive ability in both PSC-NC and PSC- EMMPRINi cells; ELISA was used to evaluate the amount of MMP2 and VEGF in media secreted by PSC- NC, PSC-EMMPRINi, as well as the co-culture cells of ASPC-NC/PSC-NC and ASPC-EMMPRINi/PSC- EMMPRINi. Results EMMPRIN mRNA was highly expressed in PSC （1 848.67 ± 281.01copies/txl）, and EMMPRIN protein mainly located in the cell membrane and cytoplasm. After silencing PSC, EMMPRIN protein expression decreased 70% -90% in PSC. There was not significant difference in the proliferative ability between PSC-NC and PSC-EMMPRINi （P = 0. 199 ）. While the invasive ability decreased significantly in PSC-EMMPRINi than in PSC-NC （P = 0. 046 ） . Conclusions Modulation of the expression of EMMPRIN in PSC by RNA interference inhibits PSC invasive ability.

关 键 词：胰腺肿瘤 RNA干扰 肿瘤侵润 细胞外基质金属蛋白酶诱导分子 胰腺星状细胞 

分 类 号：R735.9[医药卫生—肿瘤]