摘要
目的观察沉默精子相关抗原9基因(SPAG9)对裸鼠前列腺癌PC-3细胞移植瘤的抑制作用。方法利用Lipofectamine^TM2000将pGPU61绿色荧光蛋白(GFP)/Neo—SPAG9、pGPU6/GFP/Neo转染至人前列腺癌PC-3细胞。取浓度为2×10^10/L分别转染pGPU6/GFP/Neo—SPAG9和pGPU6/GFP/Neo的PC-3和PC-3细胞悬液0.2ml分别注射至各组裸鼠左腋皮下,建立前列腺癌DU-145荷瘤鼠模型,共3组,每组6只。每隔4d测量皮下移植瘤体积,绘制瘤体生长曲线。苏木素-伊红(HE)染色观察肿瘤生长。原位缺口末端标记法(TUNEL)检测移植瘤凋亡。免疫组织化学法检测各组瘤体E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、基质金属蛋白酶(MMP)-2和血管内皮生长因子(VEGF)表达。结果第27天时处死裸鼠后,转染pGPU6/GFWNeo-SPAG9组移植瘤体积为(397.9±53.1)mm^3,显著小于对照组,差异有统计学意义(P〈0.05);HE染色表明:pGPU6/GFP/Neo-SPAG9组的PC-3细胞生长抑制。TUNEL结果显示,转染pGPU6/GFP/Neo-SPAG9组平均凋亡率为(67.6±6.2)%,与对照组比较,差异有统计学意义(P〈0.05);免疫组织化学法显示,Yimentin、MMP-2、E-cadherin在转染pGPU6/GFP/Neo-SPAG9组的平均值分别为75.87±6.89、74.97±8.12、306.13±21.87,与对照组比较,差异均有统计学意义(P〈0.05)。pGPU6/GFP/Neo—SPAG9组VEGF表达量为91.49±5.29,与pGPU6/GFP/Neo组及对照组比较,差异均有统计学意义(P〈0.05)。结论沉默SPAG9可抑制前列腺癌的生长,促进其凋亡,同时减少血管的生成并降低侵袭相关蛋白的异常表达。
Objective To investigate the inhibitory effect of silencing sperm associated antigen 9 (SPAG9) gene on the transplanted prostate cancer of nude mice. Methods The pGPU6/GFP/Neo - SPAG9 and pGPU6/GFP/Neo were transfected into human prostate cancer PC - 3 cells by LipofectamineTM 2000. BALB/c nude mice were randomly divided into 3 groups, 6 in each group. The models of prostate cell carcinoma xenografts were established by subcutaneously injecting 2 ×10^7 untransfected PC - 3, pGPU6/GFP/Neo- SPAG9 transfected PC -3 and pGPU6/GFP/Neo transfected PC -3 cells into the right flank of mice. The volume of tumors was measured every 4 days, and tumor growth curve was drawn. Hematoxylin and eosin (HE) staining was used to evaluate the station of tumor cells. TdT -mediated dUTP nick end labeling (TUNEL) was used to assay the apoptosis of tumor cells. The immunohistochemistry was used to detect the expression of E -cadherin, Vimentin, matrix metalloproteinase (MMP) -2 and vascular endothelial growth factor (VEGF). Results All mice were sacrificed at the 27th day, and the final volume of tumor in pGPU6/GFP/Neo -SPAG9 group was (397. 9 ±53. 1 ) mm^3 , significantly less than in the control group (P 〈 0.05). HE staining showed that the growth of PC - 3 cells was inhibited in pGPU6/GFP/ Neo - SPAG9 group. TUNEL results showed that the average apoptosis rate of pGPU6/GFP/Neo - SPAG9 was (67. 6 ±6. 2) %, significantly different from the control group (P 〈 0. 05 ). The immunohistochemistry showed that the average absorbance (A) of Vimentin, MMP- 2 and E -cadherin in pGPU6/GFP/Neo - SPAG9 group was respectively 75.87 ±6. 89, 74.97 ±8. 12 and 306. 13 ±21.87, significantly different from that in the control group (P 〈0. 05). The average A value of VEGF were 91.49 ±5.29, significantly different from that in the control group ( P 〈 0. 05 ). Conclusion Silencing SPAG9 gene can inhibit the growth of prostate cancer, promote its apoptosis, and reduce angiogenesis and the abnormal expression of invasion related proteins.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第1期134-137,共4页
Chinese Journal of Experimental Surgery
基金
江苏省自然科学基金(BK20151166)
关键词
精子相关抗原9
前列腺癌
Silencing sperm associated antigen 9
Prostate cancer