中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2011年
4期
260-264
,共5页
曾楷峰%金海林%张伟锋%肖斌%朱宏%郝波%施瑞华
曾楷峰%金海林%張偉鋒%肖斌%硃宏%郝波%施瑞華
증해봉%금해림%장위봉%초빈%주굉%학파%시서화
食管肿瘤%胃肿瘤%血管生成拟态%缺氧诱导因子1α%RNA干扰
食管腫瘤%胃腫瘤%血管生成擬態%缺氧誘導因子1α%RNA榦擾
식관종류%위종류%혈관생성의태%결양유도인자1α%RNA간우
Esophageal neoplasms%Stomach neoplasms%Vasculogenic mimicry%Hypoxia inducible factor-1α%RNA interference
目的 探讨缺氧诱导因子1α(HIF-1α)对人食管痛及胃癌细胞体外增殖、迁移及血管生成拟态的影响.方法 将HIF-1α shRNA重组质粒pGCsi-shHIF-1 α稳定转染人食管鳞癌细胞Eca-109和胃腺癌细胞SGC-7901,采用Western blot方法检测常氧及缺氧情况下各组转染细胞HIF-1α的抑制效果,采用平板克隆形成实验和四甲基偶氮唑蓝(MTT)法检测转染细胞的增殖活性,采用Transwell小室检测转染细胞的迁移能力,采用三维培养方法观察Eca-109和SGC-7901细胞能否形成血管网状结构及转染细胞株的管腔形成能力.结果 常氧及缺氧情况下,各转染组细胞HIF-1α蛋白表达量均为0,与未转染组(Eca-109组分别为0.74±0.05和1.11±0.06,SGC-7901组分别为0.60±0.05和0.96±0.07)比较,表达均被显著抑制(均P＜0.01).与空载体组和未转染组相比,转染组细胞的增殖活性显著降低(均P＜0.05),体外迁移能力显著降低(均P＜0.01).Eca-109细胞株各组中,常氧情况下,Eca-109组和Eca-109/shRNA组形成管状结构数目分别为(30.8±3.9)个和(3.7±2.8)个,差异有统计学意义(P＜0.01);缺氧情况下,分别为(34.3±3.4)个和(3.9±2.7)个,差异有统计学意义(P＜0.01).缺氧情况下,Eca-109组形成管状结构数日较常氧时明显增加(P＜0.05).SGC-7901细胞株各组中,常氧情况下,SGC-7901组和SGC-7901/shRNA组形成管状结构数目分别为(26.2±3.4)个和(4.9±3.5)个,差异有统计学意义(P＜0.01);缺氧情况下,分别为(30.1±4.1)个和(5.3±3.6)个,差异有统计学意义(P＜0.01).缺氧情况下,SGC-7901组形成管状结构数日较常氧时明显增加(P＜0.05).结论 Eca-109细胞和SGC-7901细胞均能形成血管生成拟态管状结构.RNA于扰沉默HIF-1α能有效抑制Eca-109细胞和SGC-7901细胞增殖、迁移及体外血管生成拟态管状结构形成.
目的 探討缺氧誘導因子1α(HIF-1α)對人食管痛及胃癌細胞體外增殖、遷移及血管生成擬態的影響.方法 將HIF-1α shRNA重組質粒pGCsi-shHIF-1 α穩定轉染人食管鱗癌細胞Eca-109和胃腺癌細胞SGC-7901,採用Western blot方法檢測常氧及缺氧情況下各組轉染細胞HIF-1α的抑製效果,採用平闆剋隆形成實驗和四甲基偶氮唑藍(MTT)法檢測轉染細胞的增殖活性,採用Transwell小室檢測轉染細胞的遷移能力,採用三維培養方法觀察Eca-109和SGC-7901細胞能否形成血管網狀結構及轉染細胞株的管腔形成能力.結果 常氧及缺氧情況下,各轉染組細胞HIF-1α蛋白錶達量均為0,與未轉染組(Eca-109組分彆為0.74±0.05和1.11±0.06,SGC-7901組分彆為0.60±0.05和0.96±0.07)比較,錶達均被顯著抑製(均P＜0.01).與空載體組和未轉染組相比,轉染組細胞的增殖活性顯著降低(均P＜0.05),體外遷移能力顯著降低(均P＜0.01).Eca-109細胞株各組中,常氧情況下,Eca-109組和Eca-109/shRNA組形成管狀結構數目分彆為(30.8±3.9)箇和(3.7±2.8)箇,差異有統計學意義(P＜0.01);缺氧情況下,分彆為(34.3±3.4)箇和(3.9±2.7)箇,差異有統計學意義(P＜0.01).缺氧情況下,Eca-109組形成管狀結構數日較常氧時明顯增加(P＜0.05).SGC-7901細胞株各組中,常氧情況下,SGC-7901組和SGC-7901/shRNA組形成管狀結構數目分彆為(26.2±3.4)箇和(4.9±3.5)箇,差異有統計學意義(P＜0.01);缺氧情況下,分彆為(30.1±4.1)箇和(5.3±3.6)箇,差異有統計學意義(P＜0.01).缺氧情況下,SGC-7901組形成管狀結構數日較常氧時明顯增加(P＜0.05).結論 Eca-109細胞和SGC-7901細胞均能形成血管生成擬態管狀結構.RNA于擾沉默HIF-1α能有效抑製Eca-109細胞和SGC-7901細胞增殖、遷移及體外血管生成擬態管狀結構形成.
목적 탐토결양유도인자1α(HIF-1α)대인식관통급위암세포체외증식、천이급혈관생성의태적영향.방법 장HIF-1α shRNA중조질립pGCsi-shHIF-1 α은정전염인식관린암세포Eca-109화위선암세포SGC-7901,채용Western blot방법검측상양급결양정황하각조전염세포HIF-1α적억제효과,채용평판극륭형성실험화사갑기우담서람(MTT)법검측전염세포적증식활성,채용Transwell소실검측전염세포적천이능력,채용삼유배양방법관찰Eca-109화SGC-7901세포능부형성혈관망상결구급전염세포주적관강형성능력.결과 상양급결양정황하,각전염조세포HIF-1α단백표체량균위0,여미전염조(Eca-109조분별위0.74±0.05화1.11±0.06,SGC-7901조분별위0.60±0.05화0.96±0.07)비교,표체균피현저억제(균P＜0.01).여공재체조화미전염조상비,전염조세포적증식활성현저강저(균P＜0.05),체외천이능력현저강저(균P＜0.01).Eca-109세포주각조중,상양정황하,Eca-109조화Eca-109/shRNA조형성관상결구수목분별위(30.8±3.9)개화(3.7±2.8)개,차이유통계학의의(P＜0.01);결양정황하,분별위(34.3±3.4)개화(3.9±2.7)개,차이유통계학의의(P＜0.01).결양정황하,Eca-109조형성관상결구수일교상양시명현증가(P＜0.05).SGC-7901세포주각조중,상양정황하,SGC-7901조화SGC-7901/shRNA조형성관상결구수목분별위(26.2±3.4)개화(4.9±3.5)개,차이유통계학의의(P＜0.01);결양정황하,분별위(30.1±4.1)개화(5.3±3.6)개,차이유통계학의의(P＜0.01).결양정황하,SGC-7901조형성관상결구수일교상양시명현증가(P＜0.05).결론 Eca-109세포화SGC-7901세포균능형성혈관생성의태관상결구.RNA우우침묵HIF-1α능유효억제Eca-109세포화SGC-7901세포증식、천이급체외혈관생성의태관상결구형성.
Objective To investigate the effect of hypoxia inducible factor-1α (HIF-1α) on the proliferation, migration and vasculogenic mimicry(VM ) in human esophageal squamous cell carcinoma cell line Eca-109 and gastric adenocarcinoma cell line SGC-7901 in vitro.Methods The recombinant plasmid pGCsi-shHIF-1α was transfected into Eca-109 and SGC-7901 cells by LipofectamineTM 2000.The inhibitory effect of HIF-1α was measured at protein level by Western blot under normoxia and hypoxia.The cell proliferation was detected by colony formation and MTT assays.The migration of transfected cells was assayed using Transwell chambers.Whether Eca-109 and SGC-7901 cells could form the capillary tube-like structures (TLSs) was observed by 3-dimensional culture, and the tube formation of transfected cells was detected by tube-like structure formation assay.Results The expression of HIF-1α protein in each group of transfected cells was significantly suppressed under normoxia and hypoxia ( Eca-109: 0.00, 0.74 ± 0.05;0.00, 1.11±0.06; SGC-7901: 0.00, 0.60 ±0.05; 0.00, 0.96 ±0.07, P＜0.01).Colony formation and MTT assays showed that the cell proliferation of the pGCsi-shHIF-1α transfected cells was slower than that of the control groups (104.7±9.6, 151.7±4.5; 88.3±5.1, 128.3±6.7, P＜0.05).The migration of the recombinant plasmid-transfected cells was significantly suppressed compared with that of cells transfected with empty vector (55.5 ± 11.2, 121.9 ± 17.3; 64.7 ± 10.8, 132.3 ± 16.0, P ＜ 0.01 ).Both the Eca-109 and SGC-7901 cells could form TLSs when cultured on matrigel, and the number of tubules was significantly increased under hypoxia (30.8 ± 3.9, 34.3 ± 3.4;26.2±3.4,30.1±4.1,P＜0.05),the tubule-forming ability of transfected groups was significantly inhibited under normoxia and hypoxia ( Eca109: 3.7±2.8, 30.8±3.9; 3.9 ±2.7, 34.3 ±3.4; SGC-7901: 4.9 ±3.5, 26.2 ±3.4; 5.3 ±3.6,30.1 ±4.1, P＜0.01 ).Conclusions Both the esophageal squamous cell carcinoma cell line Eca-109 and gastric adenocarcinoma cell line SGC-7901 are capable of forming vasculogenic mimicry structures in vitro.The recombinant plasmid pGCsi-shHIF-1α can efficiently suppress their proliferation, migration and vasculogenic mimicry formation.
