摘要
目的白细胞介素-9(IL-9)是机体重要的免疫因子,参与不同肿瘤的发生发展,而其在胰腺癌的生物学效应尚不清楚,为此探索IL-9对人胰腺癌裸鼠移植瘤的作用及其机制。方法应用人胰腺癌细胞株PANC-1建立裸鼠皮下移植瘤模型。应用随机数字表法将64只裸鼠平均分成4组(16只/组),对照组予瘤内注射生理盐水0.1mL,低剂量组、中剂量组和高剂量组分别予瘤内注射90(低剂量组)、180(中剂量组)和360ng(高剂量组)IL-9,各组均隔天干预1次,共注射8次。末次用药3d后,各组随机处死8只裸鼠,取各组移植瘤组织并测质量。采用免疫组织化学染色法检测瘤组织中STAT3、p-STAT3、Cyclin D1和Bcl-2蛋白表达。蛋白质印迹法检测瘤组织中STAT3、p-STAT3。其余小鼠观察生存时间。结果低剂量组、中剂量组和高剂量组移植瘤的平均质量分别为(216.01±37.15)、(261.63±39.85)和(323.12±46.71)mg,明显高于对照组移植瘤的(196.75±25.32)mg,F=17.447,P<0.001;中(t=3.410,P=0.002)、高剂量组(t=6.643,P<0.001)与对照组相比差异有统计学意义。与对照组相比,IL-9干预后裸鼠的生存期缩短(χ~2=18.850,P=0.003),高剂量组的平均生存时间显著降低(χ~2=11.347,P=0.001)。蛋白质印迹法和免疫组化染色法检测结果显示,与对照组相比,IL-9干预后瘤组织内STAT3表达水平未见明显改变,而p-STAT3与表达水平显著升高,差异有统计学意义,均P<0.001。免疫组织化学染色结果还显示,在不同剂量IL-9干预后的瘤组织中,与对照组相比Bcl-2表达无明显变化(χ~2=1.067,P=0.956),Cyclin D1表达升高,组间差异有统计学意义,χ~2=10.416,P=0.018。结论IL-9能明显促进人胰腺癌裸鼠移植瘤生长,同时降低裸鼠的生存时间,该作用可能是通过激活STAT3磷酸化实现的。
OBJECTIVE The ain of this study was to investigate the effect and mechanism of interleukin-9 （IL-9） on human pancreatic cancer xenograft in nude mice. METHODS Human pancreatic cancer PANC-1 cells were inoculated subcutaneously into BALB/c（nu/nu） mice. When tumors reached 50-- 100 mm3 , 64 mice were randomly divided into 4 groups （n= 16） with injection of saline and different dose of IL-9 （90 ng, 180 ng, 360 ng） respectively every other day for 16 days. The tumor diameter was measured every 3 days. Eight mice of each group were sacrificed randomly on 3th day after the last treatment to measure the weight of tumor. The expression of STAT3 and p-STAT3 were detected by immu- nohistochemistry and Western blotting. Cyclin D1 and Bcl-2 expression was assessed with immunohistochemistry and quantitated. The survival time of the rest mice were recorded and compared. RESULTS The mean weight of tumors inthe control group, the low-dose group, the moderate-dose and the high-dose group were （196. 75 ± 25. 32） rag, （216.01±37.15） rag, （261.63±39.85） mg and （323.12±46.71） rag, respectively, and increased in a dose-dependent manner （F= 17. 447, P〈0. 001）. The weight of tumor in the moderate-dose and high-dose group was the most obviously higher as compared to the control group （t=3. 410,P=0. 002vs6. 643,P〈0. 001）. The average survival time of the high-dose group was （33.3 ±4. 9）d, which was significantly shortened compared with the other groups （z= 11. 347, P〈0. 001）. The result of Western blotting and immunohistochemical staining showed that after being treated with differ- ent doses of IL-9, the expression of p-STAT3 in tumor tissue was significantly higher than that of the control group （F= 13. 323, P=0. 000; Zz = 13. 867, P= 0. 003）. The expression of STAT3 in all IL-9-treated groups have no significant difference compared with the contro[ group （F= 2. 521, P = 0. 078 ; X2 = 0. 998, P=1. 000）. Immunohistochemical stai- ning also showed that treatment of tumor tissues with different doses of IL-9 increased Cyclin D1 expression（zz = 10. 416, P=0. 018）, while Bcl-2 expression have no significant changes（zz = 1. 067,P=0. 956）. CONCLUSIONS IL-9 can signif- icantly promote the growth of subcutaneous transplanted tumor derived from human pancreatic cancer PANC-1 cells and reduce the survival time of the nude mice. The mechanism might be related to activation of STAT3 pathway.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2017年第18期1268-1274,共7页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(31560257
81660107)
