[Effects of Omi/HtrA2 on expression of anti-apoptotic protein PED/PEA-15 and apoptosis of prostate cancer cell line PC-3].
OBJECTIVE
The interaction between pro-apoptotic factors and anti-apoptotic factors is closely related to the genesis and development of tumors. Omi/HtrA2 is a novel gene involved in the regulation of apoptosis. PED/PEA-15 is a widely expressed anti-apoptotic protein. This study was to explore the effects of Omi/HtrA2 on PED/PEA-15 expression and apoptosis of prostate cancer cell line PC-3.


METHODS
Omi/HtrA2 expression and specific siRNA vectors were constructed and transiently transfected into PC-3 cells. The effect of Omi/HtrA2 on PED/PEA-15 expression was assayed by Western blot, and its effect on apoptosis of PC-3 cells was analyzed by ELISA. Caspase-8 activity was assayed using Caspase-8 colorimetric assay kit. The effects of Omi/HtrA2-specific siRNA sequence on its transcription and translation were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The sensitivity of PC-3 cells to cisplatin (DDP) after Omi/HtrA2 gene silencing was determined by flow cytometry.


RESULTS
Enzyme digestion analysis and DNA sequencing confirmed Omi/HtrA2 expression, and specific siRNA vectors were successfully constructed. After transfection of Omi/HtrA2 expression vector, PED/PEA-15 expression was inhibited, Caspase-8 activity was promoted, and the apoptosis of PC-3 cells was enhanced. The sensitivity of PC-3 cells to DDP was suppressed after Omi/HtrA2 gene silencing.


CONCLUSIONS
Omi/HtrA2 can promote the apoptosis of PC-3 cells through inhibiting PED/PEA-15 expression.
Hu
Xiao-Yong
XY
Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430022, P. R. China.
Chen
Xiao-Chun
XC
Zhu
Zhao-Hui
ZH
Chen
Chao-Hui
CH
Zeng
Fu-Qing
FQ
Lu
Gong-Cheng
GC
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology; D020868:Gene Silencing; D005822:Genetic Vectors; D000074923:High-Temperature Requirement A Serine Peptidase 2; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008297:Male; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D010750:Phosphoproteins / Q000378:metabolism*; D010957:Plasmids; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D034741:RNA, Small Interfering / Q000235:genetics*; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014162:Transfection
D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D047908:Intracellular Signaling Peptides and Proteins; D024101:Mitochondrial Proteins; C092039:PEA15 protein, human; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D012697:Serine Endopeptidases; C000615914:HTRA2 protein, human; D000074923:High-Temperature Requirement A Serine Peptidase 2; D053181:Caspase 8; D002945:Cisplatin