解放军医学院学报
解放軍醫學院學報
해방군의학원학보
Academic Journal of Chinese Pla Medical School
2015年
10期
1033-1038
,共6页
高蕊%冯帆%贾辉%张帆%王涛%董国福%马德宾%马宏达%韩雅玲%刘蕾
高蕊%馮帆%賈輝%張帆%王濤%董國福%馬德賓%馬宏達%韓雅玲%劉蕾
고예%풍범%가휘%장범%왕도%동국복%마덕빈%마굉체%한아령%류뢰
miRNA153 沉默%肺癌细胞%电离辐射
miRNA153 沉默%肺癌細胞%電離輻射
miRNA153 침묵%폐암세포%전리복사
miRNA153 silencing%lung cancer cells%ionizing radiation
目的：探讨反义核酸沉默 microRNA 153(miRNA153)对电离辐射杀伤肺癌细胞株的影响。方法利用脂质体转染miRNA153的反义核酸；使用60Co-γ射线照射肺癌细胞；使用 CCK-8实验、软琼脂成集落实验(锚定非依赖性生长)和Trans-well 实验检测 miRNA153反义核酸对电离辐射杀伤肺癌细胞的影响；RT-PCR 和 Western blot 实验检测 miRNA153的反义核酸对 miRNA153、其靶标蛋白 PTEN 以及细胞存活/凋亡耐受调控蛋白 Survivin 表达的影响。结果 CCK-8实验结果显示，中等剂量射线(4 Gy)照射能够杀伤肺癌细胞 A549、H460、H1299和 H358，下调 miRNA153的表达能够增强辐射对肺癌细胞的杀伤作用；软琼脂成集落和 Transwell 实验进一步证实 miRNA153的反义核酸能够上调辐射对 A549细胞锚定非依赖性生长和侵袭的抑制作用。分子机制实验结果表明，miRNA153的反义核酸能够显著降低 miRNA153的表达水平，提高肿瘤抑制因子 PTEN 并降低细胞存活因子 Survivin 的表达。结论降低 miRNA153的表达能够上调辐射对肺癌细胞系的体外杀伤作用。
目的：探討反義覈痠沉默 microRNA 153(miRNA153)對電離輻射殺傷肺癌細胞株的影響。方法利用脂質體轉染miRNA153的反義覈痠；使用60Co-γ射線照射肺癌細胞；使用 CCK-8實驗、軟瓊脂成集落實驗(錨定非依賴性生長)和Trans-well 實驗檢測 miRNA153反義覈痠對電離輻射殺傷肺癌細胞的影響；RT-PCR 和 Western blot 實驗檢測 miRNA153的反義覈痠對 miRNA153、其靶標蛋白 PTEN 以及細胞存活/凋亡耐受調控蛋白 Survivin 錶達的影響。結果 CCK-8實驗結果顯示，中等劑量射線(4 Gy)照射能夠殺傷肺癌細胞 A549、H460、H1299和 H358，下調 miRNA153的錶達能夠增彊輻射對肺癌細胞的殺傷作用；軟瓊脂成集落和 Transwell 實驗進一步證實 miRNA153的反義覈痠能夠上調輻射對 A549細胞錨定非依賴性生長和侵襲的抑製作用。分子機製實驗結果錶明，miRNA153的反義覈痠能夠顯著降低 miRNA153的錶達水平，提高腫瘤抑製因子 PTEN 併降低細胞存活因子 Survivin 的錶達。結論降低 miRNA153的錶達能夠上調輻射對肺癌細胞繫的體外殺傷作用。
목적：탐토반의핵산침묵 microRNA 153(miRNA153)대전리복사살상폐암세포주적영향。방법이용지질체전염miRNA153적반의핵산；사용60Co-γ사선조사폐암세포；사용 CCK-8실험、연경지성집락실험(묘정비의뢰성생장)화Trans-well 실험검측 miRNA153반의핵산대전리복사살상폐암세포적영향；RT-PCR 화 Western blot 실험검측 miRNA153적반의핵산대 miRNA153、기파표단백 PTEN 이급세포존활/조망내수조공단백 Survivin 표체적영향。결과 CCK-8실험결과현시，중등제량사선(4 Gy)조사능구살상폐암세포 A549、H460、H1299화 H358，하조 miRNA153적표체능구증강복사대폐암세포적살상작용；연경지성집락화 Transwell 실험진일보증실 miRNA153적반의핵산능구상조복사대 A549세포묘정비의뢰성생장화침습적억제작용。분자궤제실험결과표명，miRNA153적반의핵산능구현저강저 miRNA153적표체수평，제고종류억제인자 PTEN 병강저세포존활인자 Survivin 적표체。결론강저 miRNA153적표체능구상조복사대폐암세포계적체외살상작용。
Objective To declare whether silencing of miRNA153 via its anti-sense nuclear (inhibitor) induces the radio-sensitization of lung cancer cells. Methods The anti-sense nuclear acid (inhibitor) of miRNA153 was transfected into lung cancer cells, which were irradiated by 60Co-γ. The CCK-8 analysis, soft-agar and Transwell assays were performed to identify the effect of miRNA153 silencing on radio-sensitization in lung cancer cells. RT-PCR and Western blot assays were used to detect the effect of anti-sense nuclear acid (inhibitor) of miRNA153 on miRNA153, its target protein PTEN and the expression of Survivin cell proliferation of survival regulators. Results CCK-8 analysis revealed that the irradiation of medium dose ray (4 Gy) could kill the lung cancer cells (A549, H460, H1299 and H358), and the down-regulation of expression of miRNA153 could enhance the radio-sensitization in lung cancer cells. The soft-agar and Transwell assays proved that the up-regulation of rays could inhibit the anchorage-independent growth and invasion of A549 cells. And the molecular mechanism experiment indicated that the anti-sense nuclear acid of miRNA153 significantly disrupted the endogenous expression of miRNA153 and Survivin, and in turn upregulated the expression of PTEN. Conclusion Silencing of miR-153 significantly enhances the sensitivity of lung cancers.
