中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2012年
6期
481-486
,共6页
喉肿瘤%癌,鳞状细胞%细胞凋亡
喉腫瘤%癌,鱗狀細胞%細胞凋亡
후종류%암,린상세포%세포조망
Laryngeal neoplasms%Carcinoma,squamous cell%Cell apoptosis
目的 观察下调组蛋白去乙酰化酶6(histone deacetylation 6,HDAC6)对人喉鳞状细胞癌(简称鳞癌)Hep-2细胞裸鼠移植瘤的抑制作用并探讨其体内抗肿瘤机制.方法 将喉鳞癌Hep-2细胞注射入裸鼠皮下,1周后裸鼠荷瘤模型建成.将裸鼠分为3组进行治疗,即空白对照组、空载体组和HDAC6小干扰RNA( siRNA)组,监测肿瘤生长变化.采用免疫组织化学法检测不同组别裸鼠移植瘤瘤体内Ki-67增殖情况的变化.采用Western blot法检测转染HDAC6前后不同组别裸鼠移植瘤组织凋亡相关基因B淋巴细胞-2(bcl-2)及bcl相关X蛋白(bcl-associated x protein,Bax)表达的变化.采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL法)检测转染HDAC6前后裸鼠移植瘤组织凋亡情况的变化.结果 HDAC6 siRNA转染荷瘤裸鼠后,与空载体组及空白对照组相比,HDAC6 siRNA组荷瘤裸鼠肿瘤体积显著下降(F =64.783,P＜0.05)；原位杂交、免疫组织化学及Western blot结果表明,HDAC6 siRNA转染组裸鼠肿瘤组织中HDAC6 mRNA及蛋白表达水平均显著下调(P值均＜0.05)；Western blot结果显示HDAC6 siRNA转染组裸鼠肿瘤组织中Bax蛋白表达显著上调而Bcl-2的表达则显著下调,三组间比较差异具有统计学意义(P值均＜0.05)；免疫组织化学结果显示转染HDAC6 siRNA组Ki-67染色阳性细胞数显著少于空载体组及空白对照组(F=25.793,P＜0.05)；TUNEL结果表明,HDAC6转染组中细胞明显发生凋亡.结论 HDAC6 siRNA能有效抑制人喉鳞癌Hep-2细胞裸鼠移植瘤的生长,并降低HDAC6和Bcl-2的表达,提高Bax的表达,为喉鳞癌的分子治疗提供了理论依据.
目的 觀察下調組蛋白去乙酰化酶6(histone deacetylation 6,HDAC6)對人喉鱗狀細胞癌(簡稱鱗癌)Hep-2細胞裸鼠移植瘤的抑製作用併探討其體內抗腫瘤機製.方法 將喉鱗癌Hep-2細胞註射入裸鼠皮下,1週後裸鼠荷瘤模型建成.將裸鼠分為3組進行治療,即空白對照組、空載體組和HDAC6小榦擾RNA( siRNA)組,鑑測腫瘤生長變化.採用免疫組織化學法檢測不同組彆裸鼠移植瘤瘤體內Ki-67增殖情況的變化.採用Western blot法檢測轉染HDAC6前後不同組彆裸鼠移植瘤組織凋亡相關基因B淋巴細胞-2(bcl-2)及bcl相關X蛋白(bcl-associated x protein,Bax)錶達的變化.採用末耑脫氧覈苷痠轉移酶介導的dUTP缺口末耑標記測定法(TUNEL法)檢測轉染HDAC6前後裸鼠移植瘤組織凋亡情況的變化.結果 HDAC6 siRNA轉染荷瘤裸鼠後,與空載體組及空白對照組相比,HDAC6 siRNA組荷瘤裸鼠腫瘤體積顯著下降(F =64.783,P＜0.05)；原位雜交、免疫組織化學及Western blot結果錶明,HDAC6 siRNA轉染組裸鼠腫瘤組織中HDAC6 mRNA及蛋白錶達水平均顯著下調(P值均＜0.05)；Western blot結果顯示HDAC6 siRNA轉染組裸鼠腫瘤組織中Bax蛋白錶達顯著上調而Bcl-2的錶達則顯著下調,三組間比較差異具有統計學意義(P值均＜0.05)；免疫組織化學結果顯示轉染HDAC6 siRNA組Ki-67染色暘性細胞數顯著少于空載體組及空白對照組(F=25.793,P＜0.05)；TUNEL結果錶明,HDAC6轉染組中細胞明顯髮生凋亡.結論 HDAC6 siRNA能有效抑製人喉鱗癌Hep-2細胞裸鼠移植瘤的生長,併降低HDAC6和Bcl-2的錶達,提高Bax的錶達,為喉鱗癌的分子治療提供瞭理論依據.
목적 관찰하조조단백거을선화매6(histone deacetylation 6,HDAC6)대인후린상세포암(간칭린암)Hep-2세포라서이식류적억제작용병탐토기체내항종류궤제.방법 장후린암Hep-2세포주사입라서피하,1주후라서하류모형건성.장라서분위3조진행치료,즉공백대조조、공재체조화HDAC6소간우RNA( siRNA)조,감측종류생장변화.채용면역조직화학법검측불동조별라서이식류류체내Ki-67증식정황적변화.채용Western blot법검측전염HDAC6전후불동조별라서이식류조직조망상관기인B림파세포-2(bcl-2)급bcl상관X단백(bcl-associated x protein,Bax)표체적변화.채용말단탈양핵감산전이매개도적dUTP결구말단표기측정법(TUNEL법)검측전염HDAC6전후라서이식류조직조망정황적변화.결과 HDAC6 siRNA전염하류라서후,여공재체조급공백대조조상비,HDAC6 siRNA조하류라서종류체적현저하강(F =64.783,P＜0.05)；원위잡교、면역조직화학급Western blot결과표명,HDAC6 siRNA전염조라서종류조직중HDAC6 mRNA급단백표체수평균현저하조(P치균＜0.05)；Western blot결과현시HDAC6 siRNA전염조라서종류조직중Bax단백표체현저상조이Bcl-2적표체칙현저하조,삼조간비교차이구유통계학의의(P치균＜0.05)；면역조직화학결과현시전염HDAC6 siRNA조Ki-67염색양성세포수현저소우공재체조급공백대조조(F=25.793,P＜0.05)；TUNEL결과표명,HDAC6전염조중세포명현발생조망.결론 HDAC6 siRNA능유효억제인후린암Hep-2세포라서이식류적생장,병강저HDAC6화Bcl-2적표체,제고Bax적표체,위후린암적분자치료제공료이론의거.
Objective To study the effect of histone deacetylation 6(HDAC6) siRNA on the growth of xenografted human laryngeal squamous cell carcinoma cell line Hep-2 in nude mice and underlying mechanism.Methods Laryngeal squamous cell carcinoma cell line Hep-2 cells were subcutaneously injected to the back of nude mice and transplanted tumor model was established after one week.Nude mice was divided into three groups including blank control group,empty vector group and HDAC6 siRNA group,and the tumor growth was observed. Ki-67 proliferation index was detected by immunohistochemistry.Western blot,in situ hybridization and immunohistochemistry were used to detect the mRNA and protein expressions of HDAC6 in xenograft.The expressions of Bcl-2 and Bax proteins were examined by Western blotting.Cell apoptosis was detected by TUNEL.Results The mean volume of xenograft transfected with HDAC6 siRNA was less than that of xenograft transfected with empty vector or that of xenograft with blank control treatment (P ＜ 0.05 ).HDAC6 siRNA effectively down-regulated the expressions of HDAC6 mRNA and the expressions of HDAC6 and Bcl-2 proteins,but up-regulated the expression of Bcl-2 protein in xenografts,with significant differences ( all P ＜ 0.05 ).The proliferation index of Ki-67 in HDAC6 siRNA transfection group was significantly lower than that in blank control group or emptry vector group ( P ＜ 0.05).TUNEL assay demonstrated that HDAC6 evidently evoked cell apoptosis( P ＜ 0.05 ).Conclusion HDAC6 siRNA could effectively inhibited the growth of xenografted human laryngeal carcinoma cell line Hep-2 in nude mice,down-regulate the expressions of HDAC6 and bcl-2,and up-regulate the expression of bax.
