Reverse transcription-polymerase chain reaction and western blotting analysis for detection of p63 isoforms in uterine cervical cancers
Reverse transcription-polymerase chain reaction and western blotting analysis for detection of p63 isoforms in uterine cervical cancers
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DOI：
10.1111/j.1525-1438.2006.00638.x
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发表时间：
2006-07-01
影响因子：
4.8
通讯作者：
Kim, I.
中科院分区：
文献类型：
--
作者：
Lin, Z.;Nan, Y.;Kim, I.
本研究的目的是探讨TAp 63和Delta Np 63亚型在宫颈癌中的作用。在7个SiHa、ME-180、SNU 17、SNU 902、SNU 1160、SNU 703和SNU 1299人乳头瘤病毒（HPV）阳性子宫颈鳞状细胞癌（SCC）细胞系，1个HT 3 HPV阴性SCC细胞系，采用逆转录-聚合酶链反应（RT-PCR）和蛋白质印迹法对一种HeLa腺癌细胞系进行了检测。新鲜非肿瘤和肿瘤组织的子宫颈和子宫内膜癌也进行了研究。宫颈癌细胞系中TA和Δ N型以及三种亚型的RT-PCR显示TA和Δ N型以及所有三种亚型的阳性条带，但HPV阴性HT 3 SCC细胞系中α亚型的弱条带，HeLa腺癌细胞系中β亚型无条带。组织标本中TA、Delta N和p63三种亚型的RT-PCR均为阳性条带，除γ亚型外，非肿瘤组织中的表达较肿瘤组织弱或不表达。在蛋白质印迹法中，癌细胞系以及非肿瘤和肿瘤组织样品均显示TA和Δ N的表达，并且仅在癌细胞系和肿瘤组织中发现γ同种型但β同种型表达（有或没有α同种型）。β亚型，可能是Delta Np 63，可能被认为是子宫颈鳞状细胞癌发生中的重要亚型。
The aim of this study was to investigate the role of TAp63 and Delta Np63 isoforms in uterine cervical cancers. The messenger RNA (mRNA) and protein expressions of TA and Delta N forms as well as alpha, beta, and gamma isoforms of p63 were studied in seven SiHa, ME-180, SNU17, SNU902, SNU1160, SNU703, and SNU1299 human papillomavirus (HPV)-positive uterine cervical squamous cell carcinoma (SCC) cell lines, one HT3 HPV-negative SCC cell line, and one HeLa adenocarcinoma cell line using reverse transcription-polymerase chain reaction (RT-PCR) and western blotting. Fresh nonneoplastic and neoplastic tissues of uterine cervical and endometrial cancers were also studied. RT-PCR for TA and Delta N form and three isoforms of p63 showed positive bands for both TA and Delta N forms and for all three isoforms in cervical cancer cell lines but weak band for alpha isoform in HPV-negative HT3 SCC cell line and no band for beta isoform in HeLa adenocarcinoma cell line. RT-PCR for TA and Delta N and three isoforms of p63 mRNA in tissue samples showed positive bands in almost all samples, except for gamma isoform, the expression was weak or absent in nonneoplastic tissues compared with neoplastic tissues. In western blotting, cancer cell lines and both nonneoplastic and neoplastic tissue samples showed expression of TA and Delta N, and gamma isoform but beta isoform expression with or without alpha isoform was only found in cancer cell lines and neoplastic tissues. beta isoform, possibly of Delta Np63, may be considered as an important isoform in uterine cervical squamous cell carcinogenesis.