RARβ基因联合全反式维甲酸抗口腔鳞癌体内研究

The antitumor effect of RARβgene combined with ATRA on human oral squamous cell carcinoma Xenografts in Athymic nude mice

目的 :观察维甲酸受体 β(RetinoicAcidReceptorbeta ,RARβ)基因联合全反式维甲酸 (All TransRetinoicAcid ,ATRA)对荷口腔鳞癌裸鼠体内抗肿瘤的作用和安全性。方法 :pSG5RARβ质粒转化、扩增、酶切鉴定。将 1× 10 7/ml密度的Tca8113细胞悬液 0 .2ml,分别接种于 2 0只裸鼠双侧臀部皮下。 2周后 ,随机将其分为对照组 ( 6只 )、ATRA治疗组 ( 8只 )、RARβ +ATRA治疗组 ( 6只 )。于接种后第 2、3周末 2次瘤内注射DNA 脂质体复合物 ,每只注射总量 6.75 μg。第 2周末开始 ,ATRA组和转RARβ基因组管饲给予 15mg/kg剂量的ATRA ,1次 /d ,5d/周× 4周。每 4d观察记录肿瘤最大径 (a)和最大垂直相交径 (b) ,观察记录动物一般情况。给药 4周后处死动物 ,切取肿瘤、称重。结果 :治疗结束时 ,对照组、ATRA、RARβ +ATRA治疗组肿瘤平均体积分别为 3 111.2mm3 ,14 2 8.8mm3 ,65 3 .9mm3 。治疗后 ,各组肿瘤平均体积分别增加 75 .1倍、66.1倍和 2 1.9倍 ,RARβ +ATRA治疗组与对照组差异有显著性 (P =0 .0 479)。 3组裸鼠平均瘤重分别为 2 .79g、0 .91g和 0 .5 7g ,ATRA和RARβ +ATRA治疗组与对照组相比差异有统计学意义 (P <0 .0 5 )。ATRA治疗组抑瘤率为 67.4% ,RARβ +ATRA治疗组抑瘤率为 79.6%。

Objective:The purpose of this study was to determine the antitumor effect of Retinoic acid receptor-beta (RARβ)gene and All trans retinioc acid (ATRA) on human oral squamous carcinoma cell xenograft in athymic nude mice.Method: The pSG5RARβ plasmids were extracted from bacteria using plasmid mini extraction kit after the proliferation of the transfected bacteria. The purified plasmid DNA was digested and confirmed using 0.8% agarose electrophoresis. Athymic nude mice (6～7 weeks old, about 20 g in weight) were implanted with 2 ×10 6 cells bilaterally into the buttocks .After two weeks, the animals were divided randomly into three groups: control group, ATRA group and ATRA +RARβ group. There were 6,8 and 6 mice in each group, respectively. The compounds of lipofectamine-DNA were injected into the tumor at each point. The same volume of normal saline was injected into the tumors of mice in control group and ATRA group. At the same time, ATRA were administered p.o to mice in ATRA and ATRA +RARβ group with a 20 gauge intragastric feeding tube daily ,5 days per week, in 0.1 ml of super-refined and sterilized sesame oil at a dose of 15 mg/kg/d for 4 weeks. The sizes of the tumors were recorded every 4 days. At the end of the treatment, all the animals were killed. Tumors were surgically excised and weighed. Result: At the end of the treatment, the average volumes of each group were 3111.2 mm 3, 1428.8 mm 3 and 653.9 mm 3, respectively. The Volumes of each group increased 75.1, 66.1 and 21.9 times, respectively. The average weights of the tumors for each group were 2.79 g, 0.91 g and 0.57 g, respectively. SAS 6.02 version statistical precedure was used for the statistic analyse. The difference of the average tumor weights between ATRA group, RARβ+ATRA group and control group was significant (P<0.05). But the difference between the two treated groups had no significance. The percentages of growth inhibition for ATRA and RARβ+ATRA group were 67.4% and 79.6%.Conclusion: Transfecting RARβ gene in vivo and administrating ATRA may be a promising stratigy for treatment of oral squamous cell carcinoma.