Change of the cell cycle after flutamide treatment in prostate cancer cells and its molecular mechanism
Change of the cell cycle after flutamide treatment in prostate cancer cells and its molecular mechanism
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DOI：
10.1111/j.1745-7262.2005.00031.x
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发表时间：
2005-12-01
影响因子：
2.9
通讯作者：
Shao, GX
中科院分区：
文献类型：
--
作者：
Wang, Y;Shao, C;Shao, GX
目的：探讨雄激素受体（AR）对前列腺癌细胞LNCaP细胞周期相关基因CDKN1A、BTG1表达的影响。方法：经AR拮抗剂氟他胺治疗后，通过相差显微镜和流式细胞术证实其作用，利用cDNA芯片分析细胞周期相关基因的差异表达。以氟他胺处理后的细胞为实验组，LNCaP细胞为对照组，分别用Cy5和Cy3染料对实验组和对照组的cDNA探针进行反转录标记。然后将cDNA探针与含有8 126个人类独特cDNA序列的cDNA芯片杂交，并对芯片进行扫描，得到每个位点上Cy5和Cy3的荧光值。初步分析后，进行逆转录聚合酶链反应（RTPCR）试验，对芯片结果进行验证。结果：经AR拮抗剂氟他胺治疗后，326个基因（3.93%）表达差异，其中下调97个，上调219个。其中，CDC10、NRAS、BTG1、Wee1、CLK3、DKFZP564A122、CDKN1A、BTG2 8个上调基因可能与细胞周期相关。RT-PCR证实实验组CDKN1A和BTG1基因mRNA表达升高，而p53 mRNA表达无明显变化。结论：氟他胺可上调前列腺癌细胞中CDKN1A和BTG1的表达。CDKN1A和BTG1蛋白表达在抑制癌细胞增殖中发挥重要作用。CDKN1A对前列腺癌细胞的细胞周期有很大影响，可能通过p53不依赖途径在癌细胞中发挥作用。前列腺癌细胞可能通过激活AR影响细胞周期相关基因，从而打破细胞周期控制。
Aim: To explore the effect of androgen receptor (AR) on the expression of the cell cycle-related genes, such as CDKN1A and BTG1, in prostate cancer cell line LNCaP. Methods: After AR antagonist flutamide treatment and confirmation of its effect by phase contrast microscope and flow cytometry, the differential expression of the cell cycle-related genes was analyzed by a cDNA microarray. The flutamide treated cells were set as the experimental group and the LNCaP cells as the control We labeled cDNA probes of the experimental group and control group with Cy5 and Cy3 dyes, respectively, through reverse transcription. Then we hybridized the cDNA probes with cDNA microarrays, which contained 8 126 unique human cDNA sequences and the chip was scanned to get the fluorescent values of Cy5 and Cy3 on each spot. After primary analysis, reverse transcription polymerase chain reaction (RTPCR) tests were carried out to confirm the results of the chips. Results:After AR antagonist flutamide treatment, three hundred and twenty-six genes (3.93%) expressed differentially, 97 down-regulated and 219 up-regulated. Among them, eight up-regulated genes might be cell cycle-related, namely CDC10, NRAS, BTG1, Wee1, CLK3, DKFZP564A122, CDKN1A and BTG2. The CDKN1A and BTG1 gene mRNA expression was confirmed to be higher in the experimental group by RT-PCR, while p53 mRNA expression had no significant changes. Conclusion: Flutamide treatment might up-regulate CDKN1A and BTG1 expression in prostate cancer cells. The protein expressions of CDKN1A and BTG1 play an important role in inhibiting the proliferation of cancer cells. CDKN1A has a great impact on the cell cycle of prostate cancer cells and may play a role in the cancer cells in a p53-independent pathway. The prostate cancer cells might affect the cell cycle-related genes by activating AR and thus break the cell cycle control.