沉默精子相关抗原9基因对裸鼠前列腺癌PC-3细胞移植瘤的抑制作用

Inhibitory effect of silencing silencing sperm associated antigen 9 gene on the transplanted prostate cancer of nude mice

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.