Munirajan A K, Kannan K, Bhuvarahamurthy V, Ishida I, Fujinaga K, Tsuchida N, Shanmugam G
Cancer Biology Division, School of Biological Sciences, Madurai Kamaraj University, India.
Gynecol Oncol. 1998 Jun;69(3):205-9. doi: 10.1006/gyno.1998.4991.
Infection with the high-risk strain of human papillomaviruses (HPVs) and the inactivation of the tumor suppressor gene p53 through mutation are important factors in cervical carcinogenesis. To know whether such events would occur in cervical carcinomas of Indians, 43 tumors (consisting of 36 of stage III B and 6 of stage II B) were screened for p53 and p16 gene mutations.
PCR followed by single-strand conformation polymorphism (SSCP) analysis were used to detect mutations in p53 and p16 genes and PCR for the presence of human papillomavirus genome. HPV status was ascertained by PCR amplification of parts of E6 and E7 genes using primers pU-1M and pU-2R and typing was carried out by restriction analysis.
Of the 43 samples analyzed, 4 samples (9%) showed mobility shifts for p53 mutations; PCR products of the p16 gene did not show band shifts in SSCP analysis. HPV DNA was detected in 70% of the 43 samples analyzed: HPV 16 in 23 cases (53%), HPV 18 in 4 cases (13.3%), and HPV 33 in 1 case (3.3%). Two amplified HPV DNAs that were difficult to type with various restriction enzymes were cloned and the amplified regions were sequenced. One of these was 93% close to HPV 35 and the other was 80% close to HPV 58. Three samples had both p53 mutations and HPV genome.
Our results indicate that HPV 16 infection was more common than HPV 18, the p53 mutations and HPV infection were not mutually exclusive events in the genesis of carcinoma of uterine cervix among Indian women, and p16 gene may not play a role in Indian cervical carcinomas.
感染高危型人乳头瘤病毒(HPV)以及通过突变使肿瘤抑制基因p53失活是宫颈癌发生的重要因素。为了解这些事件是否会在印度人的宫颈癌中发生,对43例肿瘤(其中36例为III B期,6例为II B期)进行了p53和p16基因突变筛查。
采用聚合酶链反应(PCR)继之以单链构象多态性(SSCP)分析来检测p53和p16基因的突变,并通过PCR检测人乳头瘤病毒基因组的存在情况。HPV状态通过使用引物pU - 1M和pU - 2R对E6和E7基因的部分片段进行PCR扩增来确定,分型则通过限制性分析进行。
在分析的43个样本中,4个样本(9%)显示p53突变有迁移率改变;p16基因的PCR产物在SSCP分析中未显示条带迁移。在所分析的43个样本中,70%检测到HPV DNA:23例(53%)为HPV 16,4例(13.3%)为HPV 18,1例(3.3%)为HPV 33。对两种用各种限制性酶难以分型的扩增HPV DNA进行克隆并对扩增区域进行测序。其中一个与HPV 35有93%的相似性,另一个与HPV 58有80%的相似性。3个样本同时存在p53突变和HPV基因组。
我们的结果表明,HPV 16感染比HPV 18更常见,在印度女性宫颈癌发生过程中,p53突变和HPV感染并非相互排斥的事件,并且p16基因可能在印度宫颈癌中不起作用。