[Effects of inhibiting nucleosomal binding protein 1 on proliferation of human prostate cancer cells].
OBJECTIVE
To explore the role of nucleosomal binding protein 1 (NSBP1) in the proliferation of human hormone-dependent prostate cancer cells by inhibiting its mRNA expression.


METHODS
Four well-designed short-hairpin RNA (shRNA) targeting NSBP1, including a negative control shRNA, were synthesized and inserted into the pSilencer 2.1-U6 neo plasmid. The pSilencer-81 plasmid was identified as the most efficient. The 4 recombinant plasmids, pSilencer-58, 81, 126, and Neg, were then transfected to human hormone-dependent prostate cancer cells of the line LNCaP, i.e., LNCap/81, LNCap/58, LNCap/126, and LNCap/Neg cells. RT-PCR and Western blotting were used to detect the inhibitory efficiency of different plasmids. MTT method was used to detect cell viability and flow cytometry was used to observe cell cycle distribution.


RESULTS
Western blotting showed that the protein expression of NSBP1 in the LNCap/81 cells was lower by 85% compared to that in the LNCap/Neg cells. Compared with the control cells, the A values of the LNCap/81 cells 60, 84, 108, and 132 hours after transfection were all significantly lower than those of the LNCap/Neg cells (t = 4.501, 4.282, 5.229, and 4.759, all P < 0.05), showing that the cell viability was reduced. The proportions of cells in the G(2)M and S phases 60, 84, and 108 hours after transfection of the LNCap/81 cells were all significantly lower than those of the LNCap/Neg cells (t = 3.705, 3.887, and 8.220, all P < 0.05).


CONCLUSIONS
The suppressed expression of NSBP1 in prostate cancer cells mediated by shRNA inhibits cell proliferation significantly, which indicates that NSBP1 may play an important role in the proliferation of prostate cancer cells.
Zhou
Li-Qun
LQ
Department of Urology, Peking University First Hospital, Institute of Urology, Peking University, Beijing 100034, China. zhoulqmail@china.com
Song
Gang
G
He
Zhi-Song
ZS
Hao
Jin-Rui
JR
Na
Yan-Qun
YQ
D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D024001:HMGN Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008297:Male; D010957:Plasmids / Q000235:genetics; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology; D034741:RNA, Small Interfering / Q000235:genetics*; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D024001:HMGN Proteins; C429289:HMGN5 protein, human; D034741:RNA, Small Interfering; D015534:Trans-Activators