长春新碱对INS-1细胞hTERT、Sp1、c-myc基因表达影响的研究

目的观察抗肿瘤药物长春新碱（Vincristine,VCR）对大鼠胰岛B细胞瘤株INS-1细胞增殖和侵袭的抑制作用和hTERT、Sp1、c-myc表达的影响及其意义,探讨胰岛B细胞瘤的发病机制。方法 INS-1分为对照组（不加VCR）和实验组（加入浓度分别为0.008 mg/mL、0.04 mg/mL、0.2 mg/mL、1 mg/mL的VCR）。两组培养24 h后,用MTT比色法检测INS-1细胞的增殖抑制率;用Transwell侵袭试验检测INS-1细胞的侵袭力;RT-PCR检测各组细胞中hTERT、Sp1、c-myc基因的表达。结果实验组抑制率明显高于对照组（P〈0.05）;且随着VCR浓度的增加,INS-1细胞生长抑制率逐渐增加（P〈0.05）;Transwell小室培养细胞24 h后,实验组侵袭到下层膜的细胞数为（63.26±5.12）个,对照组为（150.65±4.02）个（P〈0.01）;与对照组比较,随着时间的延长,实验组的hTERT、Sp1、c-myc mRNA基因在INS-1细胞的表达也逐渐减弱（P均〈0.05）。结论 VCR能抑制INS-1细胞增殖和侵袭,可能通过抑制Sp1、c-myc和hTERT表达,从而抑制INS-1的端粒酶活性,进而抑制细胞增殖和降低侵袭能力。

Influence of VCR on the expression of hTERT,Sp1 and c-myc in INS-1 cells

Objective To observe the effect and significance of the antitumor drugs(Vincristine,VCR) on the proliferation and invasion of rat islet beta tumor cell lines(INS-1 cell),and the expression of hTERT,Sp1and c-myc,and to explore the mechanism of Pancreatic B cell tumor.Methods The INS-1 cells were divided into two groups:the control group(no VCR) and the experimental group(concentration of VCR were 0.008 mg/ml,0.04 mg/ml,0.2 mg/ml,1 mg/ml,respectively).After being cultured for 24 hours,MTT assay was used to examine the inhibition rate of the INS-1 proliferation;the invasive ability was tested by Transwell invasion assay.The expressions of hTERT,Sp1 and c-myc in INS-1 cells were detected by RT-PCR.Results With the concentration of antitumor drugs VCR increased,the growth inhibition rate of INS-1 also increased gradually(P<0.05),and the inhibition rate of the experimental group was significantly higher than that of the control group(P<0.05);after being cultured by Transwell for 24 hours,the cell number into the submucosa membrane was(150.65±4.02) in the control group,while(63.26±5.12) in the experimental group(P<0.01);Compared with the control group,as time went by,the expressions of hTERT,Sp1,c-myc mRNA gene in the INS-1 cells of the experimental group weakened gradually(P<0.05).Conclusion VCR can inhibit the proliferation and invasion of INS-1 cells mainly through inhibiting the expressions of Sp1,c-myc and Htert,and the telomerase activity of INS-1,and then inhibiting cell proliferation and reducing the invasive ability.