小柴胡汤对鼻咽癌细胞生长的抑制作用

Study on effect of Sho-saiko-to compound on growth of nasopharyngeal carcinoma cells in CNE-bearing nude mice

目的:探讨复方小柴胡汤对鼻咽癌高分化鳞癌细胞株1(nasopharyngeal carcinoma cell line,CNE-1)和鼻咽癌低分化鳞癌细胞株2(CNE-2)生长的抑制作用,试图为该药物的临床开发与应用提供实验数据。方法:按浓度梯度配置复方小柴胡汤,用MTT实验检测不同浓度复方小柴胡汤对CNE-1/2细胞生长的影响,同时运用软琼脂克隆形成实验检测其对CNE-1/2细胞增殖能力的影响;在BALB/cA-nu裸鼠右背后皮下接种对数生长期细胞悬液(5×105个/mL)0.2 mL,成功复制肿瘤模型后随机分为5组,每组8只,于次日复方小柴胡汤按0.5,0.25,0.125 g.mL-1高、中、低3个剂量给药,同时设阳性对照组和阴性对照组,连续给药15 d后,测量肿瘤的体积和质量,计算体积抑制率和瘤重抑制率,研究复方小柴胡汤对CNE-2细胞裸鼠移植瘤的抑制作用。结果:MTT法和克隆形成实验结果提示,与阴性对照组相比,不同浓度的复方小柴胡汤处理CNE-1和CNE-2细胞24,48,72 h,发现对两株的细胞生长和增殖能力均存在不同程度的抑制作用,并随药物浓度和作用时间的增加而增强,两者结果显示其半数抑制浓度约为2.5 g.L-1;与阴性对照组比较,裸鼠体内抑瘤实验发现给复方小柴胡汤治疗组裸鼠肿瘤生长速度明显减慢,阴性对照组(0.76±0.28)g,(962.88±245.96)mm3和复方小柴胡汤低浓度治疗组(0.88±0.40)g,(1 239.66±421.93)mm3肿瘤生长明显快于高、中浓度治疗组(0.22±0.14)g,(239.31±137.07)mm3;(0.20±0.16)g,(263.42±166.57)mm3及CTX阳性对照组(0.20±0.10)g,(246.72±194.6)mm3(P<0.05)。结论:复方小柴胡汤能抑制鼻咽癌CNE-1和CNE-2细胞的生长、增殖,对CNE-2裸鼠移植瘤的生长有抑制作用。

Objective： To investigate the role of sho-saiko-to compound （SSTC） on the growth of the well-differentiated squamous cell line 1 of nasopharyngeal carcinoma （ CNE-1 ） and well-differentiated CNE-2 in tumor-bearing nude mouse, and try to supply scientific data for its clinical development. Method： SSTC were prepared by concentration gradients, and the effect of SSTC on the growth and proliferation of the CNE-1 and CNE-2 were investigated by MTT assay and soft-agar colony formation test. After setting up the subcutaneous tumor-bearing nude mouse model at the right lower back （0. 2 mL CNE-2 cell suspension, 5 × 10^5/mL）, we randomly divided forty mice into 5 groups and gave high, middle and low concentration groups of SSTC （ 0. 5, 0. 25, 0. 125 g · mL^-1 ） by intragastric administration. Positive and negative groups were set up for comparison. After constant administration for 15 days, the volume and weight of the tumor were measured for inhibition rate, so as to investigate the role of SSTC on the CNE-2 bearing tumor. Re- sult： In vitro, compared with negative control, SSTC at different gradient concentrations were cultured with the CNE-1 and CNE-2 for 24 h, 48 h and 72 h. It showed that the growth and proliferation of both cell lines were inhibited to some extent. The inhibition rate was increased as the concentration and culture time increasing. Both MTr assay and sofl-agar colony formation test showed that the 50% inhibiting concentration （ IC50 ） was about 2. 5 g · L^-1. In vivo, compared with negative control, the SSTC could slow down the tumor growth in the SSTC treated groups. The tumor growth of the negative control group （0. 76 ±0. 28） g, （962. 88 ±245.96） mm^3 and the low concentration group of SSTC （0. 88 ±0. 40） g, （ 1 239. 66 ±421.93 ） mm^3 were obviously faster than those of the high, middle concentration group of SSTC （0. 22 ±0. 14） g, （239. 31 ± 137. 07） mm^3; （0. 20 ±0. 16） g, （ 263.42 ± 166. 57） mm^3 and CTX positive control group （0. 20 ±0. 10） g, （246. 72±194. 6） mm^3（P 〈0. 05）. Conclusion： SSTC could efficiently inhibit the growth and proliferation of CNE-1 and CNE-2 in vitro, and slow down the tumor growth of the CNE-2 bearing nude mice. It may be a new compound of Chinese medicine for nasopharyngeal carcinoma therapy.