[Effects of small interfering RNA specific for Her-2 gene on biological behavior of ovarian carcinoma cell].
OBJECTIVE
To explore the effects of small interfering RNA (siRNA) specific for Her-2 gene on biological behavior of ovarian carcinoma cell.


METHODS
Her-2 siRNA recombinant plasmid and negative control plasmid were transfected into packing cell line PT67 by liposome, and PT67 was selected by puromycin later. SKOV3 was infected by the virus supernatant of stably transfected PT67 cell lines, and the stably transfected SKOV3 cell lines (SKOV3/siRNA, SKOV3/siRNA-negative) established by selection with puromycin were investigated in terms of the reduction levels of Her-2 mRNA and p185 by RT-PCR and immunohistochemistry. Cell proliferation was assayed with methyl thiazolyl tetrazolium, and cell cycle distribution and cell apoptosis were assayed with flow cytometry. The tumor growth of the null mice was analyzed after injection of SKOV3/siRNA and SKOV3/siRNA-negative into the skin.


RESULTS
(1) The stable SKOV3 cell lines with a persistent silence of Her-2 gene were established. (2) The percentages of SKOV3/siRNA in G(0)/G(1) phase and S phase were 68.6%, 15.1% respectively; while the percentages of SKOV3/siRNA-negative in G(0)/G(1) phase and S phase were 55.8%, 23.3%. (3) The percentage of SKOV3/siRNA in early apoptosis was (10.500 +/- 0.250)%, while the percentage of SKOV3/siRNA-negative was (0.340 +/- 0.010)% (P < 0.01). (4) Compared with SKOV3/siRNA-negative, the proliferation of SKOV3/siRNA was delayed obviously (P < 0.05), and the growth of the corresponding implanted tumor slowed down significantly (P < 0.01).


CONCLUSIONS
siRNA can inhibit the expression of Her-2 gene effectively, which restrains the biological behavior of ovarian carcinoma cell.
Zhou
Ying
Y
Anhui Province Key Laboratory of Molecular Medicine, Hefei 230001, China.
Ling
Bin
B
Feng
Ding-Qing
DQ
Cheng
Zhi-Xiang
ZX
Shen
Guo-Dong
GD
Gao
Ting
T
Shi
Yong-Yun
YY
Wang
Mei-Mei
MM
Zhao
Wei-Dong
WD
Zhu
Huai-Ping
HP
D000818:Animals; D017209:Apoptosis; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018734:Genes, erbB-2; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009366:Neoplasm Seeding; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis; D014162:Transfection
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018719:Erb-b2 Receptor Tyrosine Kinases