CAJ | 학술논문

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,Western blot法检测EMMPRIN在PSC中封闭效果,MTT、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EMMPRIN mRNA呈高表达(1 848.67±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浸润能力.
목적 탐토RNA간우세포외기질금속단백매유도분자(extracellular matrix metalloproteinase inducer, EMMPRIN)적표체대이선성상세포(pancreatic stellate cell,PSC)증식、침습급전이능력적영향. 방법 응용QRT-PCR법급세포면역조화법검측EMMPRIN재PSC중mRNA함량급단백적표체;PSC경RNA간우후,Western blot법검측EMMPRIN재PSC중봉폐효과,MTT、연경지법검측세포증식능력,Transwell법검측EMMPRIN봉폐전후PSC(PSC-NC,PSC-EMMPRINi)세포침습능력,ELISA법검측MMPRIN봉폐전후PSC(PSC-NC, PSC-EMMPRINi)급여ASPC세포연합배양후(ASPC-NC/PSC-NC,ASPC-EMMPRINi/PSC-EMMPRINi)배양기중MMP2、VEGF적분비량. 결과 PSC중EMMPRIN mRNA정고표체(1 848.67±281.01) copies/μl,EMMPRIN단백주요정위우PSC적세포막급세포질중,PSC경RNA간우후,EMMPRIN단백재PSC중하조70％～ 90％,EMMPRIN봉폐전후PSC증식능력차이무통계학의의(P =0.199),EMMPRIN봉폐후PSC-EMMPRINi급연합배양ASPC-EMMPRINi/PSC-EMMPRINi적배양기중MMP2분비량교봉폐전PSC-NC급ASPC-NC/PSC-NC초감소(P=0.16),PSC-EMMPRINi교PSC-NC침습능력현저강저(P=0.046). 결론 간우EMMPRIN재PSC중적표체가억제기침습침윤능력.
Objective To investigate the role of extracellular matrix metalloproteinase inducer molecule (EMMPRIN) on proliferative, invasive and metastatic ability of pancreatic stellate cells (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/μl),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.