Shen Lu-Ying, Lin Cong-Meng
Department of Hematology, Zhangzhou Hospital Affiliated to Fujian Medical University, Zhangzhou 363000, Fujian Province, China.
Department of Hematology, Zhangzhou Hospital Affiliated to Fujian Medical University, Zhangzhou 363000, Fujian Province, China,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Jun;27(3):833-838. doi: 10.19746/j.cnki.issn.1009-2137.2019.03.031.
To explore the expression level of PLK1 in mantle cell lymphoma(MCL), and the effect of silencing PLK1 gene by RNA interference on the cell proliferation, apoptosis, and cell cycle.
S-P immunohistochemistry technique was used to detect the expression of PLK1 in tissues of 42 patients with MCL and 30 patients with reactive proliferative lymphodenitis(RPL), their expression levels were compared and analyzed. The Jeko-1 cells were transfected with lentivirus contaiming PLK-1 shRNA, then the mRNA and protein expression of PLK-1 was detected by real-time guantitative PCR and Western blot nespectively, and the silencing efficacy of PLK-1 shRNA was identificd. The cell proliferation was detected by CCK method, the cell apoptosis was detected by Annexin V/PI double staining, the cell cycle was detected by PI single staining, the changes of apoptosis-related proteins BAX, BCL-2 and Caspase 3 were detected by Western blot.
The positive expression rate of PLK-1 in tissue of MCL patients was 66.67%(28/42), which was significanfly higher than 20%(6/30) in tissue of RPL patients (P<0.05). The PLK-1 positive expression correlated with B symptom, IPI score, Ann-Arbor stage(P<0.05). After infection of Jeko-1 cells with lentivirus containing PLK-1 shRNA for 72 hours, the mRNA and protein expressions of PLK-1 were significantly down-regulated(P<0.05), the proliferation rate of cells in group of PLK-1 shRNA was significanly lower than that in control and Neg shRNA groups(P<0.05); the apoptosis rate of cells in PLK-1 shRNA group was (27.42±3.44)%, which was significantly higher than that in control group (1.23±0.42)% and Neg shRNA group (2.07±0.58) % (P<0.05). The cell cycle analysis showed that the cell ratio in G/M phase of PLK-1 shRNA group was (27.21±3.59) %, which was higher than that in control group (13.28±2.63)% and Neg shRNA group (14.34±2.37) %. The detection of apoptosis-related proteins showed that the expression of BAX was up-regulated, the expression of BCL-2 was down-regnlated and the expression of caspase 3 was up-regulated.
The PLK-l overexpression appears in tissue of MCL patients. The silencing PLK-1 gene can inhibit the proliferation of Jeko-1 cells, induce the apopotosis of Jeko-1 cells and arrestes cell cycle in G/M phase.
探讨PLK1在套细胞淋巴瘤(MCL)中的表达水平,以及RNA干扰沉默PLK1基因对细胞增殖、凋亡和细胞周期的影响。
采用S-P免疫组化技术检测42例MCL患者和30例反应性增生性淋巴结炎(RPL)患者组织中PLK1的表达,比较并分析其表达水平。将携带PLK-1 shRNA的慢病毒转染Jeko-1细胞,然后分别通过实时定量PCR和Western blot检测PLK-1的mRNA和蛋白表达,鉴定PLK-1 shRNA的沉默效果。采用CCK法检测细胞增殖,Annexin V/PI双染法检测细胞凋亡,PI单染法检测细胞周期,Western blot检测凋亡相关蛋白BAX、BCL-2和Caspase 3的变化。
MCL患者组织中PLK-1的阳性表达率为66.67%(28/42),显著高于RPL患者组织中的20%(6/30)(P<0.05)。PLK-1阳性表达与B症状、国际预后指数(IPI)评分、Ann-Arbor分期相关(P<0.05)。用携带PLK-1 shRNA的慢病毒感染Jeko-1细胞72小时后,PLK-1的mRNA和蛋白表达均显著下调(P<0.05),PLK-1 shRNA组细胞的增殖率显著低于对照组和阴性对照shRNA组(P<0.05);PLK-1 shRNA组细胞的凋亡率为(27.42±3.44)%,显著高于对照组的(1.23±0.42)%和阴性对照shRNA组的(2.07±0.58)%(P<0.05)。细胞周期分析显示,PLK-1 shRNA组G/M期细胞比例为(27.21±3.59)%,高于对照组的(13.28±2.63)%和阴性对照shRNA组的(14.34±2.37)%。凋亡相关蛋白检测显示,BAX表达上调,BCL-2表达下调,caspase 3表达上调。
MCL患者组织中出现PLK-1过表达。沉默PLK-1基因可抑制Jeko-1细胞增殖,诱导Jeko-1细胞凋亡,并使细胞周期阻滞于G/M期。