新合成的紫草萘醌类衍生物TEISHNZ的细胞毒作用和诱导人鼻咽癌细胞凋亡

Cytotoxicity and apoptosis induction of new naphthoquinone-like derivative TEISHNZ on human nasopharyngeal cancer cell

目的探讨一种新的人工半合成紫草萘醌类衍生物（2，3，11-三乙巯基-6-异己萘茜，简称TEISHNZ）。在体外对8种人癌细胞的细胞毒作用及诱导人鼻咽癌细胞（CNE2）细胞凋亡的作用。方法应用噻唑蓝（MTT）比色法检测TEISHNZ对8种人癌细胞的增殖抑制作用，以半数抑制浓度（IC50）为指标进行评价。应用流式细胞术（FCM）检测TEISHNZ诱导CNE2细胞凋亡的作用及其对细胞周期的影响。结果TEISHNZ在0．78～25μg／mL质量浓度下，对8种人癌细胞均有明显的增殖抑制作用；对CNE2细胞、人肺腺癌细胞（GLC-82）、人肝癌细胞（Bel-7402）、人白血病细胞（K562）、人宫颈癌细胞（HeLa）、人胃腺癌细胞（MGC-803）、人乳腺癌细胞（MDA453）和人口腔癌细胞（KB）的IC50分别为3．96、2．48、5．13、1．92、4．54、5．88、1．35和3．44μg／mL，IC50值均在6μg／mL以下。FCM检测结果显示，以TEISHNZ（1．56、3．12和6．25μg／mL）处理CNE2细胞48h后，在FCM的DNA直方图上可见亚二倍体（亚G1峰）；细胞凋亡率依次为3．9％、10．8％和28．1％，而对照组的细胞凋亡率为2．2％；TEISHNZ对CNE2细胞的诱导凋亡作用呈浓度依赖性；细胞周期分析显示CNE。细胞被阻滞于G2／M期。用6．25μg／mL TEISHNZ处理CNE2细胞12、24、36和48h后亦可见细胞凋亡，细胞凋亡率分别为4．5％、27．2％、48．2％和24．9％，而对照组的凋亡率为1．0％。CNE2细胞被TEISHNZ处理的前36h，其凋亡率随着作用时间的延长而增加，被阻于S和G2／M期；作用48h时，被阻于G2／M期。结论TEISHNZ对多种人癌细胞有明显的细胞毒作用，且能诱导CNE2细胞凋亡，并使细胞周期阻滞于S期和（或）G2／M期。

Objective To explore the cytotoxicity on eight kinds of human cancer cell lines and apoptosis induction on human nasopharyngeal cancer cells （CNE2） of a new semi-synthesized naphthoquinone derivative （2, 3, 11-triethanesulfanyl-6-isohexenylnaphthazarin, TEISHNZ）. Methods Inhibition of TEISHNZ on eight kinds of human cancer cell lines proliferation was assayed using MTT method. The cytotoxicity was evaluated using IC50 value. Apoptosis induction and effect on cell cycle distribution of TEISHNZ on human nasopharyngeal cancer cells were assayed by Flow Cytometry （FCM）. Results Under the 0. 78-25μg/mL of concentrations, TEISHNZ had markedly proliferated inhibition on human cancer cell lines. The IC50 values of TEISHNZ on CNE2, GLC-82, Bel-7402, K562, HeLa, MGC-803, MDA453, and KB cells were 3.96, 2.48, 5.13, 1.92, 4. 54, 5.88, 1.35, and 3.44μg/mL, respectively. The IC50 values were less than 6 μg/mL. CNE2 cells were treated respectively for 48 hby TEISHNZ （1.56, 3.12, and 6.25μg/mL） and then the CNE2 cells were detected by FCM. The result showed sub-G peak, the apoptotic rates were 3.9%, 10. 8%, and 28. 1%, respectively, and apoptotic rates of control group was 2.2%, apoptosis induction of TEISHNZ on CNE2 cell was in a concentration dependent manner, cell cycle analysis indicated that CNE2 cells were blocked in G2/M phase. After treatment with TEISHNZ （6. 25μg/mL） for 12, 24, 36, and 48 h, CEN2 cell apoptotic rates were 4. 5%, 27.2%, 48.2%, and 24.9%,respectively, and apoptotic rate of control group was 1.0%. The apoptotic rates increased with longing of treatment time, before CNE2 cells of treatment for 36 h, CNE2 cells, were blocked in S and GJM phase, after treatment for 48 h, CNE2 cells blocked in G2/M phase. Conclusion TEISHNZ against various human carcinoma cells shows marked cytotoxicity and could induce cell apoptosis, and cell cycle arrest at S phase and （or） G2/M phase.