Ramirez P T, Gershenson D M, Tortolero-Luna G, Ramondetta L M, Fightmaster D, Wharton J T, Wolf J K
Department of Gynecologic Oncology, University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
Gynecol Oncol. 2001 Dec;83(3):543-8. doi: 10.1006/gyno.2001.6438.
The purpose of this study was to determine whether transfection of ovarian cancer cell lines with recombinant adenoviral vectors containing wild-type p16(INK4a), p21(WAF1/Cip-1), and p53 caused growth inhibition and induction of apoptosis. We also measured the expression of the cell-cycle mediators Bax, Bcl-2, pRb, and mdm-2.
We introduced the wild-type p16(INK4a), p21(WAF1/Cip-1), and p53 genes into the ovarian cancer cell lines SK-OV-3 (p16(INK4a) and p53 null) and OVCA-420 (p16(INK4a) and p53 wild-type) by adenoviral transfection. Cell growth inhibition was measured over a 10-day period. Induction of apoptosis was tested for both cell lines 48 h after cell transfection. Expression of cell-cycle mediators was evaluated by Western blot analysis and densitometry.
Growth inhibition was documented after transfection with p16(INK4a), p21(WAF1/Cip-1), and p53 in both SK-OV-3 cells and OVCA-420 cells. Apoptosis was greatest in SKOV-3 cells after transfection with p53. A significant expression of Bax was only seen in the SKOV-3 cells transfected with p53. The bcl-2 protein was poorly expressed in both cell lines. Expression of pRb was suppressed in OVCA-420 cells transfected with p16(INK4a) and p21(WAF1/Cip-1). Infection with Adp16(INK4a) and Adp53 led to an increase in the level of mdm-2 in the SK-OV-3 cell line only.
In the ovarian cancer cell lines studied, cell growth was inhibited after transfection with p16(INK4a), p21(WAF1/Cip-1), and p53. Cell cycle arrest was highest with p53 transfection. The expression of pro-apoptosis proteins was primarily a function of p53 expression.
本研究旨在确定用携带野生型p16(INK4a)、p21(WAF1/Cip-1)和p53的重组腺病毒载体转染卵巢癌细胞系是否会导致生长抑制和诱导凋亡。我们还检测了细胞周期调节因子Bax、Bcl-2、pRb和mdm-2的表达。
我们通过腺病毒转染将野生型p16(INK4a)、p21(WAF1/Cip-1)和p53基因导入卵巢癌细胞系SK-OV-3（p16(INK4a)和p53缺失）和OVCA-420（p16(INK4a)和p53野生型）。在10天的时间内测量细胞生长抑制情况。在细胞转染48小时后，对两种细胞系进行凋亡诱导检测。通过蛋白质印迹分析和光密度测定法评估细胞周期调节因子的表达。
用p16(INK4a)、p21(WAF1/Cip-1)和p53转染后，SK-OV-3细胞和OVCA-420细胞均出现生长抑制。用p53转染后，SKOV-3细胞中的凋亡最为明显。仅在用p53转染的SKOV-3细胞中观察到Bax的显著表达。两种细胞系中Bcl-2蛋白的表达均较低。在用p16(INK4a)和p21(WAF1/Cip-1)转染的OVCA-420细胞中，pRb的表达受到抑制。仅在SK-OV-3细胞系中，感染Adp16(INK)和Adp53导致mdm-2水平升高。
在所研究的卵巢癌细胞系中，用p16(INK4a)、p21(WAF1/Cip-1)和p53转染后细胞生长受到抑制。p53转染导致的细胞周期阻滞最为明显。促凋亡蛋白的表达主要是p53表达的作用。
