中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2015年
6期
811-816
,共6页
周海波%刘万平%周方正%陈洁%刘合波%邹传涛%张东升%吴文学%聂龙
週海波%劉萬平%週方正%陳潔%劉閤波%鄒傳濤%張東升%吳文學%聶龍
주해파%류만평%주방정%진길%류합파%추전도%장동승%오문학%섭룡
血管内皮生长因子类/药理学%RNA,小分子干扰%鼻咽肿瘤/药物疗法%细胞增殖%肿瘤转移
血管內皮生長因子類/藥理學%RNA,小分子榦擾%鼻嚥腫瘤/藥物療法%細胞增殖%腫瘤轉移
혈관내피생장인자류/약이학%RNA,소분자간우%비인종류/약물요법%세포증식%종류전이
Vascular endothelial growth factors/PD%RNA,small interfering%Nasopharyngeal neoplasms/DT%Cell proliferation%Neoplasm metastasis
目的 以特异性siRNA在转录水平下调VEFG表达,观察对人鼻咽癌细胞株(CNE2)生长、转移等生物学活性的影响并探讨其可能分子机制.方法 构建和筛选VEGF-siRNA高效干扰质粒并测序鉴定,转染研究细胞株,观察干预VEGF表达对鼻咽癌细胞增殖、细胞周期及转移的影响.结果 以siRNA转染CNE2细胞,VEGF在基因转录和蛋白水平表达明显降低,癌细胞增殖明显受抑并呈时间依赖性;干预组癌细胞平板克隆形成能力明显降低、细胞失巢凋亡率显著增加,癌细胞增殖周期发生G1期阻滞,CyclinD1及p-Erk表达降低.结论 干预VEGF基因转录可经Erk/MAPK信号通路抑制鼻咽癌细胞增殖等生物学行为,提示VEGF为潜在的鼻咽癌基因治疗的有效靶目标.
目的 以特異性siRNA在轉錄水平下調VEFG錶達,觀察對人鼻嚥癌細胞株(CNE2)生長、轉移等生物學活性的影響併探討其可能分子機製.方法 構建和篩選VEGF-siRNA高效榦擾質粒併測序鑒定,轉染研究細胞株,觀察榦預VEGF錶達對鼻嚥癌細胞增殖、細胞週期及轉移的影響.結果 以siRNA轉染CNE2細胞,VEGF在基因轉錄和蛋白水平錶達明顯降低,癌細胞增殖明顯受抑併呈時間依賴性;榦預組癌細胞平闆剋隆形成能力明顯降低、細胞失巢凋亡率顯著增加,癌細胞增殖週期髮生G1期阻滯,CyclinD1及p-Erk錶達降低.結論 榦預VEGF基因轉錄可經Erk/MAPK信號通路抑製鼻嚥癌細胞增殖等生物學行為,提示VEGF為潛在的鼻嚥癌基因治療的有效靶目標.
목적 이특이성siRNA재전록수평하조VEFG표체,관찰대인비인암세포주(CNE2)생장、전이등생물학활성적영향병탐토기가능분자궤제.방법 구건화사선VEGF-siRNA고효간우질립병측서감정,전염연구세포주,관찰간예VEGF표체대비인암세포증식、세포주기급전이적영향.결과 이siRNA전염CNE2세포,VEGF재기인전록화단백수평표체명현강저,암세포증식명현수억병정시간의뢰성;간예조암세포평판극륭형성능력명현강저、세포실소조망솔현저증가,암세포증식주기발생G1기조체,CyclinD1급p-Erk표체강저.결론 간예VEGF기인전록가경Erk/MAPK신호통로억제비인암세포증식등생물학행위,제시VEGF위잠재적비인암기인치료적유효파목표.
Objective To investigate down-regulation of vascular endothelial growth factor (VEGF) gene at mRNA level on effects of nasopharyngeal carcinoma (NPC) cell proliferation and analyze the key molecule expressions of extracellular-signal regulated protein kinase/mitogen-activated protein kinase (Erk/MAPK) pathway to explore its molecular mechanism.Methods Highest-efficiency VEGF-siRNA plasmid was successfully constructed and screened,confirmed by sequencing,and then transfected to CNE2 cell line.Interference efficiency was quantitatively evaluated by a fluorescence quantitation polymerase chain reaction assay.Expressions of VEGF,cell cycle regulator cyclin D1,and key molecules in Erk/MAPK pathway were analyzed with Western blot.Cell proliferation was tested by cell counting kit-8 (CCK-8) kit.Clone formation rate was checked with plate clone formation assay.Alteration of cell cycle was measured with flow cytometry.Stable cell clones were screened by G418.Results VEGF expression in NPC cells transfected with specific siRNA were significantly down-regulated at the levels of mRNA or protein,and cell proliferations were inhibited in a time-dependent manner,with significant decreases of colony-forming,cyclinD1,and p-Erk,and with cell cycle arrested at G1 phase.Conclusions Intervencing VEGF gene transcription inhibited NPC cell proliferation through Erk/MAPK signaling pathway,and suggesting that VEGF should be a novel therapeutic target for NPC.