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目的探讨我们自行设计的STAT3(signal transducer and activator of transcription 3)反义核酸(ASODN)对裸鼠肺腺癌A549细胞移植瘤的成瘤能力和移植瘤的抑制作用。方法将经全硫代修饰的STAT3 ASODN加入到培养的A549细胞内,CCK-8法检测细胞生长的抑制率,然后将这种细胞接种于裸鼠颈部皮下,观察肿瘤出现的时间和肿瘤体积的变化,并计算成瘤率。同时取未经处理的A549细胞接种于裸鼠颈部皮下建立移植瘤模型,待瘤结节直径大于5 mm后,分为STAT3ASODN实验组、生理盐水组和无义寡核苷酸对照组。实验组用15 mg/kg ASODN每天1次直接瘤内注射,连续3周,隔日测量肿瘤大小。处理30 d后,剥离瘤块称质量。蛋白质免疫印迹法检测肿瘤中STAT3、p-STAT3的表达。结果 STAT3 ASODN显著抑制A549细胞生长,与无义寡核苷酸组相比,差异有统计学意义(P<0.05)。STAT3 ASODN作用后的A549细胞在裸鼠皮下成瘤能力明显降低,抑瘤率达75.8%。在各组荷瘤动物模型中,生理盐水对照组和无义寡核苷酸对照组的瘤体进行性增大,而STAT3 ASODN处理组的瘤块生长缓慢,与2个对照组相比均有差异(P<0.01),处理30 d后,ASODN处理组的肿瘤质量与2个对照组相比差异均有统计学意义(P<0.05),STAT3 ASODN处理组裸鼠A549细胞移植瘤的生长明显受到抑制,抑瘤率为51.1%。肿瘤中STAT3蛋白表达水平以及磷酸化水平也明显下调。结论 STAT3 ASODN能降低A549细胞的成瘤率、抑制移植瘤生长。
OBJECTIVE: To investigate the antitumor activity of transplanted human lung adenocarcinoma A549 cells and the inhibitory effect of transplanted tumor on nude mice with adenocarcinoma A549 cell line STAT3 (ASODN). Methods STAT3 ASODN modified with all-thio was added to cultured A549 cells. The inhibitory rate of cell growth was detected by CCK-8 method. Then the cells were inoculated subcutaneously into the neck of nude mice to observe tumor appearance time and tumor volume The change and calculate the rate of tumor formation. At the same time, untreated A549 cells were inoculated into nude mice subcutaneously to establish a xenograft tumor model. When the tumor nodules were larger than 5 mm in diameter, they were divided into STAT3ASODN experimental group, normal saline group and non-sense oligonucleotide control group. Experimental group with 15 mg / kg ASODN once daily intratumoral injection, for 3 consecutive weeks, the size of the tumor was measured every other day. After 30 days of treatment, peel masses were weighed. Western blotting was used to detect the expression of STAT3 and p-STAT3 in tumor. Results STAT3 ASODN significantly inhibited the growth of A549 cells compared with non-sense oligonucleotide group, the difference was statistically significant (P <0.05). A549 cells after STAT3 ASODN treatment significantly reduced the subcutaneous tumorigenicity in nude mice, with a tumor inhibition rate of 75.8%. In each group of tumor-bearing animal model, the tumors of the control group and the non-sense oligonucleotide control group increased progressively, while the tumor growth of STAT3 ASODN group was slow compared with the two control groups (P <0.01). After treated for 30 days, the tumor mass of ASODN group was significantly different from that of the two control groups (P <0.05). The growth of the transplanted tumor of A549 cells in STAT3 ASODN group was obvious Suppressed, tumor inhibition rate was 51.1%. STAT3 protein expression in tumor and phosphorylation levels were significantly down-regulated. Conclusion STAT3 ASODN can reduce the tumorigenic rate of A549 cells and inhibit the growth of xenografts.