p16基因治疗喉鳞状细胞癌的实验研究

Adenovirus p16(INK4A) gene therapy for laryngeal squamons cell carcinoma

目的 探讨外源性p16基因对人喉鳞状细胞癌 (简称鳞癌 )的抑制作用。方法 应用携带p16基因的复制缺陷型腺病毒转染人喉鳞癌细胞株 ,用Western免疫印记法 (Westernblot法 )检测p16基因在喉鳞癌细胞中的表达情况 ,在体外、体内观察p16基因治疗喉癌细胞的生长抑制情况。结果 复制缺陷型腺病毒载体可有效地将外源性p16基因转染入人喉鳞癌细胞株中并使其表达p16蛋白 ;体外、体内抑瘤实验表明 :携带p16基因的重组腺病毒Ad p16治疗组肿瘤生长明显受抑制 ,其肿瘤体积均数为 (91 0 0± 6 3 2 )mm3,而携带LacZ基因的重组腺病毒Ad LacZ和磷酸盐缓冲液(phosphaticbufferedsaline ,PBS)组肿瘤体积均数则分别为 (13 7 0 0± 9 62 )mm3和 (144 0 0± 13 87)mm3,经统计学检验、实验组与对照组差异有显著性 (P <0 0 5 )。Ad LacZ与PBS组比较差异无显著性 (P>0 0 5 ) ,提示p16基因可抑制人喉鳞癌细胞生长 ,而腺病毒载体对肿瘤生长无影响。结论 用腺病毒载体可有效地介导肿瘤抑制基因p16的表达 ,并可抑制人喉鳞癌细胞生长 。

Objective To assess the anti tumor effects of p16 INK4A gene transfer in laryngeal squamous cell carcinoma Methods A Complete p16 INK4A gene was inserted into a replication defective recombinant adenovirus (Ad p16) and the tumor cells were infected with Ad p16 Confirmation of p16 INK4A protein expression after Ad p16 infection was performed by Western Blotting The therapeutic effects were evaluated both in in vitro and in vivo study Results The replication defective recombinant adenovirus can direct a high level of p16 INK4A protein expression in laryngeal squamous cell carcinoma Studies both in in vitro and in vivo experiments demonstrated that Ad p16 treatment significantly inhibits the cell growth and the established tumors in nude mice The mean value of tumor volumes among the Ad p16, Ad Lacz and phosphatic buffered saline(PBS) groups was (91 00±6 32)mm 3,(137 00±9 62)mm 3 and (144 00±13 87)mm 3 respectively Statistic analysis showed that there was a significant difference between the Ad p16 group and the control groups ( P <0 05) No significant difference was found between Ad lacZ and PBS groups, which indicated that the antitumor effects was not influenced by the adenovirus Conclusion These results demonstrate a significant antitumor effect of Ad p16 against human laryngeal squamous cell carcinoma This data also further support the potential application of Ad p16 to treat the human head and neck cancer