Effects of Thalidomide on the Proliferation and Gap Junction Intercellular Communication of Prostate Cancer PC-3 Cells
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摘要:
目的 观察沙利度胺对前列腺癌PC-3细胞增殖和凋亡的作用及对PC-3细胞中Cx43表达水平的影响。 方法 将处于对数生长期的前列腺癌PC-3细胞分别给予递增浓度的沙利度胺(0、12.5、25、50、100μg/mL)处理24 h和48 h。采用CCK-8法检测不同浓度沙利度胺对PC-3细胞的增殖抑制影响, Annexin V-FITC/PI双染色法检测细胞凋亡情况, RT-PCR法检测Cx43mRNA的表达及Westernblot检测Cx43蛋白的表达。 结果 沙利度胺浓度在25~100μg/mL能明显抑制PC-3细胞体外增殖, 随着时间、浓度增加, 抑制率相应升高。不同浓度组沙利度胺处理PC-3细胞24~48 h后, Cx43 mRNA和蛋白有不同程度的升高(P < 0.05)。 结论 沙利度胺可通过抑制人前列腺癌PC-3细胞增殖、诱导其凋亡, 产生抗肿瘤作用。沙利度胺可上调前列腺癌PC-3细胞Cx43mRNA及蛋白的表达水平, 并可能促进前列腺癌PC-3细胞的细胞缝隙连接通讯功能的恢复, 从而抑制肿瘤生长。 Abstract:Objective This study aimed to observe the effects of thalidomide (THD) on the proliferation and apoptosis of pros tate cancer PC-3 cells, as well on the expression of the intercellular gap junction protein Cx43 in these cells in vitro. Methods THI with different concentrations (0, 12.5, 25, 50, and 100 μg/mL) was given to PC-3 cells at the logarithmic growth phase for 24 and 48 A Cell Counting Kit-8 was used to detect the growth rate among the medication groups with different THD concentrations and times of drug administration. The apoptosis rate was studied by annexin V-fluorescein isothiocyanate/propidium iodide double staining wit flow cytometry. Reverse-transcription polymerase chain reaction was used to detect the expression of Cx43 mRNA in the PC-3 cell, Western blot analysis was used to assay the expression of Cx43 protein in the cells. Results 1) Below 25-100 μg/L, THD considerabl inhibited the proliferation of PC-3 cells in vitro. The inhibitory action was enhanced in a dose- and time-dependent manner. 2) The expression of Cx43 mRNA gene and its protein in the groups with PC-3 cells increased to different degrees in after treatment with THD various concentrations (P < 0.05). Conclusion 1) THD has an antitumor activity in depressing the proliferation of PC-3 cells, therel: inducing cell apoptosis in vitro. 2) THD can increase the expression levels of Cx43 gene mRNA and protein, and promote the functional refreshment of gap junction intercellular communication in prostate cancer PC-3 cells, thereby inhibiting tumor growth. -
Key words:
- Thalidomide /
- Prostate carcinoma /
- Gap junction intercellular communication /
- Connexin
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图 1 沙利度胺对PC-3细胞增殖的影响
Figure 1. Effects of thalidomide on the proliferation of PC-3 cells by using Cell Counting Kit-8
图 2 不同浓度沙利度胺对PC-3细胞凋亡的影响
A：空白对照组；B：1.25 μg/mL组；C：25 μg/mL组；D：50 μg/mL组；E：100 μg/mL组
Figure 2. Effect of thalidomide on the apoptosis of PC-3 cells
图 3 不同浓度沙利度胺对PC-3细胞中Cx43 mRNA表达的影响
M：Marker；1~5：递增浓度THD（0、12.5、25、50、100 μg/mL）处理PC-3细胞24 h后；6~10：递增浓度THD（0、12.5、25、50、100 μg/mL）处理PC-3细胞48 h后
Figure 3. Effect of thalidomide of different concentrations on expression of Cx43 mRNA in PC-3 cells
图 4 不同浓度沙利度胺对PC-3细胞中Cx43蛋白表达的影响
1：对照组；2：12.5 μg/mL组；3：25 μg/mL组；4：50 μg/mL组；5：100 μg/mL组
Figure 4. Effect of thalidomide at different concentrations on expression of Cx43 protein in PC-3 cells
表 1 不同浓度沙利度胺对PC-3细胞抑制率 %
Table 1. Inhibitory effects of thalidomide at different concentrations on the proliferation of PC-3 cells
表 2 不同浓度沙利度胺处理PC-3细胞48 h后的凋亡率 %
Table 2. Effect of thalidomide at different concentrations on the apoptosis of PC-3 cells (48 h after therapy)
表 3 不同浓度沙利度胺对PC-3细胞Cx43 mRNA表达的影响
Table 3. Effect of thalidomide at different concentrations on the expression of Cx43 mRNA in PC-3 cells
表 4 不同浓度沙利度胺对PC-3细胞Cx43蛋白表达的影响
Table 4. Effect of thalidomide at different concentrations on the expression of Cx43 protein in PC-3 cells
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