体外测试全反维甲酸对鼻咽癌细胞药物效应阈和毒性阈的方法

Explore a method to measure the threshold of the effective or cytotoxity of trans-retinoic acid on nasopharyngeal carcinoma cells in vitro

目的:探讨体外全反维甲酸(trans-retinoicacid,TRA)对鼻咽癌细胞药物效应阈和毒性阈及其测试方法。方法:以鼻咽癌细胞CNE系体外培养,观察等比浓度系列(10-3～10-9mol/L)的全反维甲酸作用后,比较研究鼻咽癌细胞的克隆形成率(cloningplatingefficiency,CPE)和克隆细胞倍增率(cloningcelldoublinggrowthefficiency,CDE)的变化,并探讨细胞对药物作用的效应阈和毒性阈方法。结果:鼻咽癌细胞克隆形成率与克隆细胞倍增率随药物浓度的改变而改变,即与空白对照组的比较,尽管全反维甲酸浓度小于10-6mol/L时的克隆形成率都呈现出无明显差别(P>0 05),但当全反维甲酸一旦等于或大于10-6mol/L时,克隆形成率则立即随药物浓度升高而呈现更明显的抑制(全反维甲酸等于10-6mol/L时,P<0 05;全反维甲酸等于10-5mol/L时,则P<0 01),这说明,浓度为10-6mol/L是全反维甲酸在发挥其诱导细胞分化而促使细胞克隆形成率抑制的一个药效阈值。另一方面,当全反维甲酸在等于或小于10-6mol/L作用时,克隆细胞倍增率都呈现出无明显差别(P>0 05),而当全反维甲酸等于10-5mol/L时,克隆细胞倍增率却呈现出明显的抑制(P<0 05),说明,全反维甲酸浓度为10-5mol/L又是全反维甲酸在发挥其细胞生长抑制作用,即对其细胞群体倍增显著影响的一个毒性阈值。

Aim: To explore a method to measure the thresholds of the effective or cytotoxity of trans-retinoic acid on nasopharyngeal carcinoma cells in vitro. Methods: The nasopharyngeal carcinoma cell line(CNE) was suspended and cultured in RPMI-1640 medium containing 10% fetal serum and a series ratio concentration (10^(-3)～10^(-9) mol/L) of trans-retinoic acid (TRA) to give a final concentration then comparatively studied on the cloning plating efficiency and the cell doubling growth efficiency in vitro. Results: The effects of trans-retinoic acid on the cloning plating efficiency and the cell doubling growth efficiency of nasopharyngeal carcinoma cells were changed at a different concentration of TRA: compared with that of the control group (c(TRA)=0 mol/L), the cell doubling growth efficiency which was demonstrated positively consistent correlation to the cell proliferation was inhibited insignificantly at or below a concentration of 10^(-6) mol/L(P>0.05) but significantly at a concentration of 10^(-5) mol/L (P<0.05), and the cloning plating efficiency which was demonstrated negative consistent correlation to the cell differentiation was inhibited insignificantly below a concentration of 10^(-6) mol/L (P>0.05) but significantly at or above a concentration of 10^(-6) mol/L (P<0.01). The results indicated: The inhibition of the cloning plating efficiency on nasopharyngeal carcinoma cells induced by the concentration of 10^(-6) mol/L of trans-retinoic acid may be resulted in the cell differentiation, and that was a effective threshold; On the other hands, the inhibition of the cell cloning growth efficiency on nasopharyngeal carcinoma cells induced by the concentration of 10^(-5) mol/L of trans-retinoic acid may be resulted in cytotoxity, and that was a cytotoxic threshold. Conclusion: The cytotoxic threshold and the effective threshold of trans-retinoic acid were different in the experiment. It was suggested that identifying the concentration between the (cytotoxic) threshold and the effective threshold could provide a method to the proper application of trans-retinoic acid in the clinic or the research field.