维生素C联合硼替佐米对人T细胞淋巴瘤细胞株Jurkat细胞凋亡的应用

Application of vitamin C combined with bortezomib in human T cell lymphoma cell line Jurkat cell apoptosis

目的研究维生素C联合硼替佐米对人T细胞淋巴瘤细胞株Jurkat细胞凋亡相关蛋白B淋巴细胞瘤-2(Bcl-2)与Bcl-2相关X蛋白(Bax)表达调节的影响。方法采用30nmol/L的硼替佐米组,100μg/mL的维生素C组及联合组分别作用于Jurkat细胞24、48、72h,设立未加药物的对照组,应用倒置显微镜观察细胞的生长方式,免疫组织化学检测Bax及Bcl-2的表达,CCK-8及流式细胞技术检测Jurkat细胞凋亡率和细胞周期。结果维生素C使Jurkat细胞的生长方式从对照组的局部聚集向近乎均匀分散变化;维C组及联合组在24h至48h期间,Bcl-2的表达降低较硼替佐米组快,Bax表达升高的亦快,且维C组24~48h对Bax的表达增量远超过硼替佐米组。CCK-8检测发现,在72h时维C组、联合组、硼替佐米组Jurkat细胞的抑制率分别为25.72%、75.23%、56.81%;流式检测凋亡率在72h分别为81.73%、67.06%与81.50%;维生素C组的早期凋亡率大于晚期凋亡率,而硼替佐米组与联合组均为早期凋亡率低,但维生素C的添加,联合用药组细胞的早期凋亡率得到了大约15.26%的提高。结论维生素C抑制T细胞淋巴瘤细胞株Jurkat细胞的增殖,尤其是促进Jurkat细胞早期凋亡,且细胞凋亡调控蛋白Bax与Bcl-2参与了维生素C诱导Jurkat细胞凋亡机制。

Objective To study the influence of vitamin C combined with bortezomib on the apoptosis regulatory protein B lymphocytoma-2（Bcl-2）and Bcl-2associated X protein（Bax）expression regulation in human T-cell lymphoma cell line Jurkat cells.Methods 30nmol/L bortezomib group,100μg/mL vitamin C group and the combination group were adopted to act on Jurkat cells for 24,48,72 hrespectively.There was no drug added to the control group.The inverted microscope was used to observe the cell growth way.The expression of Bax and Bcl-2proteins was detected by immunohistochemistry.CCK-8and flow cytometry were used to detect the cellular apoptosis and cellular cycle.Results Vitamin C made the Jurkat cell growth way to change from the local accumulation growth in the control group to almost evenly disperse change.The expression decrease of Bcl-2from 24 hto 48hin the vitamin C group and combination group were faster than that in the bortezomib group,the Bax expression increase was also faster,moreover the expression increment of Bax from 24 hto 48hby vitamin C far surpassed that in the bortezomib group.The inhibition rates of Jurkat cell in three groups detected by CCK-8at 72 hwere 25.72%,75.23% and 56.81%respectively.The apoptosis rates at 72 hin three groups detected by flow cytometry were 81.73%,67.06% and 81.50% respectively.Early apoptosis rate in the vitamin C group was greater than the late apoptosis rate,but the bortezomib group and combination group all had low early apoptosis rate,but cell early apoptosis rate in the combination group was increased by about 15.26%after adding vitamin C.Conclusion Vitamin C inhibits T-cell lymphoma cell line Jurkat cell proliferation,especially promotes early apoptosis of Jurkat cells,moreover confirms that the apoptosis regulatory proteins Bax and Bcl-2participate in the vitamin C-induced Jurkat cell apoptosis mechanism.