MSP58基因RNA干涉后对人脑胶质瘤细胞裸鼠移植瘤的影响

目的观察RNA干涉法（RNA interference，RNAi）抑制人脑胶质瘤U251细胞的MSP58基因表达后，对其在裸鼠体内成瘤性及增殖、侵袭性的影响。方法将特异性的MSP58干涉表达载体pSilencer3．1．MSP58转染人脑胶质瘤细胞系U251（U251．S），同时构建相应的阴性对照组U251-NC，以及空载体组U251-H1neo。运用逆转录酶-多聚酶链反应（RT—PCR）及蛋白质印迹法（Westernblot）检测各组细胞的MSP58mRNA和蛋白表达水平。用各组U251细胞建立人脑胶质瘤裸鼠皮下移植瘤模型。结果成功建立了具有稳定下调MSP58基因表达的U251-S细胞。U251-S细胞MSP58的表达无论在mRNA水平还是在蛋白质水平均较U251组、U251-H1neo组及U251-NC组细胞显著降低（P〈0．01）。与接种U251、U251-H1neo和U251-NC细胞的裸鼠相比，接种U251-S细胞的裸鼠肿瘤形成时间延迟，肿瘤生长缓慢，肿瘤体积及瘤重均明显减小（P〈0．01）。结论MSP58基因RNAi后抑制了移植瘤细胞MSP58mRNA及蛋白的表达，进而明显抑制移植瘤的增殖和侵袭能力。

The effect of MSP58 RNAi on the human brain glioma cell xenograft in nude mice

Objective To investigate the changes of the tumorigenesis, proliferation and invasion of human brain glioma cell U251 in nude mice after suppression of MSP58 by RNA interference. Methods Human glioma cells （ U251 ） were transfected with the specific siRNA expression vector （ pSilencer3. 1- MSP58） （U251-S）. A corresponding site-mutated vector （with pSilencer3.1-NC ） and empty vector （with empty vector pSilencer3. 1-Hlneo） were constructed as a negative control （U251-NC） and empty vector control （U251-Hlneo）. The expression of MSP58 mRNA and its protein among the stable transfected cells and the untransfected ones were detected by semi-quantitative RT-PCR and Western blot, respectively. U251, U251-H1 neo, U251-NC and U251-S cells were inoculated respectively in flank subcutaneous tissue of nude mice to establish xenograft models of human brain glioma. Results The stable transfected cell lines： U251-S, U251-NC and U251-Hlneo were established. The expression levels of MSP58 gene mRNA and protein were significantly lower in U251-S than in U251-NC, U25 1-Hlneo and those untransfected U251 cells （ P 〈 0. 01 ）. The tumorigenesis time delayed, tumor grew slowly, both tumor volume and tumor weight were decreased significantly （P 〈0.01 ） in U251-S group compared with those in U251, U251-Hlneo and U251-NC cells groups. Conclusion The above results suggest that the expressions of MSP58 mRNA and protein are inhibited by MSP58 gene RNA interference and accordingly are associated with the remarkable inhibition of proliferation and invasion of xenograft glioma.