Pei X F, Wu M
Department of Cell Biology, Chinese Academy of Medical Sciences, Beijing.
Cancer Lett. 1993 Oct 15;74(1-2):15-24. doi: 10.1016/0304-3835(93)90038-b.
Two human keratinocyte cell lines transformed by human papillomavirus type 16, designated Vp and Up, were compared for their clonal growth potential and clonal origin. Up showed greater anchorage-independent growth in soft agar and higher efficiency of single-cell colony formation than Vp (24.3% compared to approximately 10%). The clonal growth potential of these two cell lines was not related to the level of HPV16 gene expression. Fourteen single cell clones of the Vp and 24 of the Up were selected, propagated and analyzed by Southern and Northern blot analysis. Clonal variations existed among subclones of each cell line and between the two cell lines. These variations included cell morphology, growth potential, and expression levels of involucrin (a differentiation marker of keratinocytes) and of HPV16 mRNAs. The Vp and Up cell lines also showed different patterns of HPV16-DNA integration and RNA transcription. However, all subclones of Vp and subclones of Up displayed identical HPV16 DNA integration and RNA expression patterns. The results suggest that both cell lines were monoclonal in origin and that the host genetic factors play an essential role in determining cell clonality.
对两个人乳头瘤病毒16型转化的人角质形成细胞系（分别命名为Vp和Up）的克隆生长潜能和克隆起源进行了比较。与Vp相比，Up在软琼脂中表现出更强的非贴壁依赖性生长以及更高的单细胞集落形成效率（分别为24.3%和约10%）。这两个细胞系的克隆生长潜能与HPV16基因表达水平无关。选取了Vp的14个单细胞克隆和Up的24个单细胞克隆，进行传代培养，并通过Southern印迹和Northern印迹分析进行检测。每个细胞系的亚克隆之间以及两个细胞系之间均存在克隆变异。这些变异包括细胞形态、生长潜能、外皮蛋白（角质形成细胞的一种分化标志物）和HPV16 mRNA的表达水平。Vp和Up细胞系还表现出不同的HPV16 - DNA整合和RNA转录模式。然而，Vp的所有亚克隆和Up的亚克隆均显示出相同的HPV16 DNA整合和RNA表达模式。结果表明，这两个细胞系均起源于单克隆，并且宿主遗传因素在决定细胞克隆性方面起着至关重要的作用。
