New alternatively spliced variant of prostate-specific membrane antigen             PSM-E suppresses the proliferation, migration and invasiveness of prostate cancer             cells.
PSM-E is a newly discovered alternatively spliced variant of prostate-specific             membrane antigen (PSMA). In the current study, its role on the proliferation,             invasiveness and migration in prostate cancer cell lines was analyzed. PSM-E and             PSMA (as a comparison) eukaryotic expression vectors pcDNA3.0/PSM-E and pcDNA3.0/PSMA             were constructed, validated by RT-PCR and Western blotting, and PSMA/PSM-E overexpression             PC-3 cell models were built. Gene interference was used to block PSMA and the             expression of its splice variants in LNCap cells. Three shRNA fragments were synthesized             against PSMA, cloned into the vector pSilencer 2.1-U6-neo, their interference             effect was evaluated by RT-PCR and Western blotting, and pSilencer 2.1-U6-neo‑shRNA3             (named p‑shRNA3) was chosen in further analyses. Growth curves were drawn to observe             the proliferation change, which showed that PSM-E had the potential to suppress             proliferation (P<0.05), but no significant change was observed in PSMA/PC-3             cells and in PSMA/PSM-E interfering LNCap cells (P>0.05). Cross-river test             showed that the migration speeds of PSM-E/PC-3 and PSMA/PC-3 were both significantly             slower than the vector negative control, and faster in p-shRNA3 interfering LNCap             cells compared with its vector negative control (P<0.05), and no significant             difference existed between PSM-E/PC-3 and PSMA/PC-3 (P>0.05). Transwell assay             showed that the invasive cells of both PSMA/PC-3 and PSM-E/PC-3 were fewer compared             to the vector negative control (P<0.05), and the invasive suppression effect             of PSM-E was weaker than PSMA (P<0.05), and accordingly, invasiveness of interfering             LNCaP cells was enhanced compared with the vector negative control (P<0.05).             These results showed that PSM-E could suppress proliferation, migration and invasiveness             of prostate cancer cells. Its suppression effect on cell proliferation is stronger             compared to PSMA and the suppression effect on invasiveness is weaker than that             of PSMA.
Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Kai-Yuan
KY
Cao
Lin
L
Xu
Ding-Mei
DM
Zhang
Xiao-Ming
XM
Zhang
Tian
T
Zhang
Xia
X
He
Zhu
Z
Wang
Fa-Shen
FS
Feng
Shao-Peng
SP
Qiu
Guan-Xin
GX
Shen
D000954:Antigens, Surface / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D043425:Glutamate Carboxypeptidase II / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism
D000954:Antigens, Surface; D020033:Protein Isoforms; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II