Shiwei Huo1, Wenyan Du2, Peng Shi3, Yuexiu Si4, Shengtian Zhao5. 1. Shandong University Jinan 250100, Shandong, China ; The Central Hospital of Taian Taian 271000, Shandong, China. 2. The Branch of Tai'an Central Hospital Taian 271000, Shandong, China. 3. The Central Hospital of Taian Taian 271000, Shandong, China. 4. The Fifth Hospital of Jinan Jinnan 250100, Shandong, China. 5. The Second Hospital of Shandong University Jinan 250100, Shandong, China.
Abstract
OBJECTIVES: To investigate the role of spermatogenesis-associated protein 6 (SPATA6) in the testicular germ cell tumors (TGCTs). METHODS: Human embryonic carcinoma (EC)-derived cell line NTera2 was employed and randomly divided into normal control group, SPATA6c group, siSPATA6c group, and SPATA6c + siSPATA6c group. The recombinant expression vector pcDNA3.1 (+)-SPATA6 and target sequence for SPATA6-specific siRNA was transfected into NTera2 cells in the SPATA6c group and siSPATA6c group, respectively. The SPATA6 protein levels in each group were determined by Western blot. Cell proliferation and apoptosis rate were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2 5-diphenyl-2Htetrazolium bromide (MTT) colorimetric assay and flow cytometry (FCM) assay, respectively. In addition, Western blot was performed to investigate the expression of Bax and B-cell lymphoma (Bcl)-2 in each group. RESULTS: Compared with control group, protein levels of SPATA6 were significantly reduced in the siSPATA6c group, but were statistically increased in the SPATA6c group (P < 0.05). Similarly, the cell viability was significantly decreased by transfection with SPATA6 siRNA, but was increased by transfection with pcDNA3.1 (+)-SPATA6 compared with the control group. Moreover, the percentages of apoptosis cell were significantly higher in siSPATA6 group than those in the three groups. After transfection of SPATA6 siRNA, the expression of Bax was significantly increased, but the expression of Bcl-2 was markedly decreased than that in the control group and SPATA6c group. CONCLUSION: SPATA6 may play an important role in TGCTs, and down-regulation of SPATA6 could lead to apoptosis of TGCTs.
OBJECTIVES: To investigate the role of spermatogenesis-associated protein 6 (SPATA6) in the testicular germ cell tumors (TGCTs). METHODS:Humanembryonic carcinoma (EC)-derived cell line NTera2 was employed and randomly divided into normal control group, SPATA6c group, siSPATA6c group, and SPATA6c + siSPATA6c group. The recombinant expression vector pcDNA3.1 (+)-SPATA6 and target sequence for SPATA6-specific siRNA was transfected into NTera2 cells in the SPATA6c group and siSPATA6c group, respectively. The SPATA6 protein levels in each group were determined by Western blot. Cell proliferation and apoptosis rate were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2 5-diphenyl-2Htetrazolium bromide (MTT) colorimetric assay and flow cytometry (FCM) assay, respectively. In addition, Western blot was performed to investigate the expression of Bax and B-cell lymphoma (Bcl)-2 in each group. RESULTS: Compared with control group, protein levels of SPATA6 were significantly reduced in the siSPATA6c group, but were statistically increased in the SPATA6c group (P < 0.05). Similarly, the cell viability was significantly decreased by transfection with SPATA6 siRNA, but was increased by transfection with pcDNA3.1 (+)-SPATA6 compared with the control group. Moreover, the percentages of apoptosis cell were significantly higher in siSPATA6 group than those in the three groups. After transfection of SPATA6 siRNA, the expression of Bax was significantly increased, but the expression of Bcl-2 was markedly decreased than that in the control group and SPATA6c group. CONCLUSION:SPATA6 may play an important role in TGCTs, and down-regulation of SPATA6 could lead to apoptosis of TGCTs.
Entities:
Keywords:
Spermatogenesis-associated protein 6; apoptosis; testicular germ cell tumors
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