紫草素对鼻咽癌CNE-2细胞增殖和侵袭的影响

Effects of shikonin on proliferation and invasion in nasopharyngeal carcinoma CNE-2 cells

目的研究紫草素对鼻咽癌CNE-2细胞增殖、侵袭及磷脂酰肌醇3激酶/蛋白激酶B(PI3K/PKB)信号通路的影响。方法体外培养鼻咽癌CNE-2细胞,低、中、高剂量实验组分别以终浓度为1,5,10μg·mL-1 紫草素处理,对照组则以等量生理盐水处理,各组分别干预24,48,72 h。用活细胞计数(CCK-8)法检测鼻咽癌CNE-2细胞增殖情况;流式细胞仪检测细胞凋亡率;用Transwell小室实验检测鼻咽癌CNE-2细胞侵袭能力;用蛋白免疫印迹(Wb)法检测鼻咽癌CNE-2细胞磷酸化PI3K(p-PI3K)蛋白和磷酸化PKB(p-PKB)蛋白表达情况。结果干预72 h时,低、中、高剂量实验组鼻咽癌CNE-2细胞增殖抑制率分别为(19.13±1.19)%,(46.23±2.42)%和(85.61±4.24)%;干预72 h时,对照组和低、中、高剂量实验组鼻咽癌CNE-2细胞凋亡率分别为(5.12±1.69)%,(16.12±3.02)%,(42.32±5.17)%,(68.79±6.38)%;侵袭率分别为(95.27±3.86)%,(76.81±3.12)%,(46.42±2.98)%,(39.13±2.74)%;鼻咽癌CNE-2细胞p-PI3K蛋白相对表达量分别为0.89±0.12,0.46±0.07,0.24±0.04,0.18±0.02;p-PKB蛋白相对表达量分别为0.93±0.11,0.51±0.08,0.22±0.03,0.17±0.02。低、中、高剂量实验组鼻咽癌CNE-2细胞增殖抑制率随紫草素作用浓度增大呈逐渐升高趋势(P<0.05);与对照组相比,低、中、高剂量实验组鼻咽癌CNE-2细胞凋亡率显著升高,而侵袭率及p-PI3K和p-PKB蛋白相对表达量降低,且呈剂量依赖性(均P<0.05)。结论紫草素可有效抑制鼻咽癌CNE-2细胞增殖,诱导其凋亡,并降低其侵袭力及p-PI3K和p-PKB蛋白表达水平,其机制可能与抑制PI3K/PKB信号通路活性有关。

Objective To investigate the effects of shikonin on proliferation, invasion and phosphatidylinositol 3-kinase/protein kinase B(PI3 K/PKB) signaling pathway in nasopharyngeal carcinoma CNE-2 cells. Methods Nasopharyngeal carcinoma CNE-2 cells were cultured in vitro, and Exp-L/M/H groups were treated with 1, 5 and 10 μg·mL-1 shikonin, control group was treated with the same amount of normal saline. Each group was treated for 24, 48, 72 h. Live cell count(CCK-8) method was used to detect the proliferation of nasopharyngeal carcinoma CNE-2 cells;flow cytometry was used to detect apoptosis rate;Transwell chamber assay was used to detect invasion ability of nasopharyngeal carcinoma CNE-2 cells. The expression of phosphorylated PI3K(p-PI3K) protein and phosphorylated PKB(p-PKB) protein in nasopharyngeal carcinoma CNE-2 cells were detected by Western blotting(Wb). Results The inhibition rates of CNE-2 cell proliferation in Exp-L/M/H groups were(19. 13 ± 1. 19) %,(46. 23 ± 2. 42) % and(85. 61 ± 4. 24) %,the apoptotic rates of CNE-2 cells in nasopharyngeal carcinoma in control group and Exp-L/M/H groups were(5. 12 ± 1. 69) %,(16. 12 ± 3. 02) %,(42. 32 ± 5. 17) % and(68. 79 ± 6. 38) %;the invasive rates were(95. 27 ± 3. 86) %,(76. 81 ± 3. 12) %,(46. 42 ± 2. 98) % and(39. 13 ± 2. 74) %;the relative expression levels of p-PI3K protein in nasopharyngeal carcinoma CNE-2 cells were0. 89 ± 0. 12,0. 46 ± 0. 07,0. 24 ± 0. 04 and 0. 18 ± 0. 02,the relative expression levels of p-PKB protein were 0. 93 ± 0. 11,0. 51 ± 0. 08,0. 22 ± 0. 03 and 0. 17 ± 0. 02. The inhibition rates of CNE-2 cell proliferation in Exp-L/M/H groups were increased gradually with the increase of shikonin concentration(P < 0. 05). Compared with control group,the apoptosis rates of nasopharyngeal carcinoma CNE-2 cells in Exp-L/M/H groups were significantly increased,while the invasive rates and the relative expression of p-PI3K and p-PKB protein were decreased,and the trend was concentration-dependent(all P < 0. 05). Conclusion Shikonin can effectively inhibit the proliferation of nasopharyngeal carcinoma CNE-2 cells,induce apoptosis,and reduce its invasiveness and expression of p-PI3K and p-PKB protein,its mechanism may be related to inhibition of PI3K/PKB signaling pathway activity.