Musashi-1在非小细胞肺癌组织的表达及其对人肺腺癌细胞A549生物学行为的影响

Expression of Musashi - 1 in non - small cell lung cancer and its effects on biological behaviors of human pulmonary adenocarcinoma cell line A549

目的检测Musashi-1在非小细胞肺癌（NSCLC）组织中的表达，观察RNA干扰（RNAi）靶向下调Musashi-1基因表达对人肺腺癌A549细胞生物学功能的影响。方法应用免疫组织化学法检测54例NSCLC组织及配对癌旁组织Musashi-1蛋白表达水平，利用脂质体Lipofectamine＇rM2000RNAiMax将Musashi-1小干扰RNA（siRNA）转染至人肺腺癌细胞A549中，采用反转录．聚合酶链反应（RT—PCR）及Westernblot法分别检测Musashi-1mRNA，3-连环蛋白（3-catenin）mRNA及其蛋白的表达；噻唑蓝（MTF）法检测细胞的增殖；划痕实验及Transwell侵袭小室实验检测细胞的迁移和侵袭能力。结果54例NSCLC组织中，Musashi-1表达率为68．5％，而癌旁组织中的表达率仅为13．0％，差异有统计学意义（x2=34．51，P〈0．05）；siRNA干扰实验组Musashi—lmRNA和β-cateninmRNA的表达量分别为9．51±2．04和11．38±3．02，明显低于阴性对照组的17．51±2．13和25．08±2．91和空白对照组的16．38±2．08和26．07±3．04（P〈0．05）。siRNA干扰实验组Musashi-1和β-catenin蛋白的灰度值分别为0．401±0．015和0．304±0．021，明显低于阴性对照组（0．742±0．011和0．825±0．023）和空白对照组（0．796±0．010和0．810±0．024，P〈0．05）。生长曲线显示Musashi-1siRNA转染组细胞的增殖能力较阴性对照组及空白对照组明显减慢（P〈0．05）；同时，转染细胞的侵袭和迁移能力较对照组细胞明显降低，差异有统计学意义（P〈0．05）。结论下调Musashi-1在人肺腺癌细胞A549的表达后，细胞的增殖能力减弱，迁移及侵袭能力降低。

Objective To investigate tile expression of Musashi -1 in non -small cell lung cancer （NSCLC） and study the effect of Musashi -1 down -regulation by small interfering RNA （siRNA） on the biological features of lung cancer cell line A549. Methods The expression of Musashi - I protein in 54 NSCLC specimens and paired adjacent non - cancerous tissues were detected by immunohistochemistry. Musashi- 1 siRNA was transfected into A549 cells with LipofectamineTM 2000 RNA interference （RNAi） Max. The expression levels of Musashi - 1 and β - eatenin were detected by using reverse transcription pol- ymerase chain reaction （RT- PCR） and Western blotting respectively after transfection. The proliferation of A549 cells was measured by methyl thiazol tetrazolium （MTT） assay. Cell migration and invasion were assessed by using wound - healing assay and Transwell chambers respectively. Results In 54 patients with NSCLC, the expression rate of Musashi - 1 protein was higher in cancerous tissue （68.5%） compared with that in paired non - cancerous tissues （ 13.0% ）, and the difference between the two groups was sta- tistically significant （X2 = 34. 51, P 〈 0. 05 ）. The expression of Musashi - 1 mRNA and β - catenin mRNA in the Musashi- 1 siRNA transfected group were 9. 51 ±2.04 and 11.38 ± 3.02 respectively. As compared with the negative control group （ 17.51 ± 2. 13 and 25.08 ± 2. 91 ） and non - transfected group （ 16. 38 ± 2. 08 and 26. 07 ± 3.0g）, the difference was significant （ P 〈 0. 05 ）. The protein expression of Musashi - 1 and β - catenin in the Musashi - 1 siRNA transfected group were 0. 401 ±0. 015 and 0. 304 ± 0. 021 respectively. As compared with the negative control group （0. 742 ±0. 011 and 0. 825 ±0. 023） and non - transfected group （0. 796± 0. 010 and 0. 810 -± 0. 024）, the difference was significant （ P 〈 0. 05 ）. The curves of cell growth indicated that cell proliferation in the Musashi - 1 siRNA group was inhibited sig- nificantly. The migration and invasion capacity of A549 cells were also effectively inhibited after Musashi - 1 i knockdown. Conclusion Musashi - 1 may be related with proliferation, migration and invasion of A549 cells, suggesting that Musashi - 1 is a potential therapeutic target for lunz cancer.