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).
Conclusion: siRNA can inhibit the expression of Her-2 gene effectively, which restrains the biological behavior of ovarian carcinoma cell.
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