人乳头瘤病毒16 E6和E7基因对人喉鳞癌细胞系增殖和分化的影响

目的研究人乳头瘤病毒（human papillomavirus，HPV）16E6和E7基因对Lscc-02人喉鳞癌细胞系增殖和分化的影响，以探讨HPV16与喉鳞癌演进的关系。方法采用阳离子脂质体基因转染技术将携带有HPV16和E7DNA的真核表达质粒导入HPV阴性的Lscc-02喉鳞癌细胞系，以载体质粒转染的细胞以及未转染细胞为对照，观察并比较体外及裸鼠体内细胞的增殖状态和分化程度。结果转染HPV16和E7的喉鳞癌细胞体外增殖加快，增殖A比值为2．96，高于载体质粒转染的细胞1．90以及未转染细胞1．85，软琼脂克隆形成率较载体质粒转染的细胞以及未转染细胞增大，分别为23．6％、12．7％、12．0％，血清依赖降低；增殖核抗原Ki-67表达的阳性百分数为93．8％，明显高于载体质粒转染细胞的80．7％以及未转染细胞的79．2％；角蛋白13表达的阳性百分数为80．9％，明显低于载体质粒转染细胞的91．0％以及未转染细胞的93．7％；裸鼠体内转染HPV16和E7的喉鳞癌细胞移植瘤潜伏期及体内倍增时间较载体质粒转染的细胞以及未转染细胞缩短，潜伏期分别为19d、28d、30d，体内倍增时间分别为2．15d、3．28d,3．47d，瘤组织细胞变小，异形性增大，细胞核分裂象多，有向低分化鳞癌发展趋势。结论HPV16和E7促进Lscc-02喉鳞癌细胞系增殖，抑制其分化，使肿瘤细胞的恶性增强，提示HPV16在喉鳞癌的演进过程中可能起重要作用。

Effects of human papillomavirus 16 E6 and E7 oncogene on the proliferation and differentiation of human laryngeal squamous cell carcinoma cell line

Objective To investigate the correlation between HPV16 and the development of laryngeal squamous cell carcinoma by studing the effects of HPV16 E6 and E7 oncogene on the proliferation and differentiation of human laryngeal squamous carcinoma cell line. Methods Cationic phosphonolipid was used to transfect pLXSN16E6E7 into Lscc-02 high differentiation laryngeal squamous cell carcinoma cell line, the cells transfected by the vector pLXSN and Lscc-02 cells served as control groups. The changes of proliferation and differentiation were measured in vitro and in vivo. Results The proliferation was quicken in experimental group. The average A ratio reached to 2.96, which was higher than 1.90 and 1.85 of control groups. The clone forming rate was 23. 6% in experimental group, which was higher than 12. 7% and 12. 0% of control groups. Serum-dependant became lower in experimental group. The Ki-67 positive expression rate was markedly increased and cytokeratin 13 positive expression rate was markedly decreased in experimental group as compared with control groups. The Ki-67 positive expression rates were 93. 8%, 80. 7% and 79. 2% respectively. The cytokeratin 13 positive expression rate were 80. 9%, 91.0% and 93. 7% respectively. The latent period and internal double time of transplant tumor were short in experimental group compared with control groups. The latent period were 19 d, 28 d and 30 d respectively. The internal double time of transplant tumor was 2. 15 d, 3. 28 d and 3.47 d respectively. The volume of transplant tumor was smaller in the same period in experimental group. The cells of transplant tumor in experimental group showed an image of small size, great allotype and a trend of low differentiation. Conclusions HPV16 E6 and E7 can promote the proliferation and inhibit the differentiation of Lscc-02laryngeal squamous cell carcinoma cells which suggest HPVI6 may play an important rule in the development of laryngeal squamous cell carcinoma.