Objective: To observe the effects of short hairpin RNA (shRNA) targeting Her2 on its gene expression when the shRNA was stably transfected into human ovarian cell lines, SKOV3 and SKOV3.ip1, which have different extent of malignancy and investigate the changes of the biological characters of the two cell lines after the stable transfection.
Methods: The plasmids expressing shRNA targeting Her2 gene were transfected into SKOV3 and SKOV3.ip1 cells. The stably transfected cells were gained by antibiotic screening. The expression of Her2 before and after the transfection was detected by RT-PCR and western blot. The transwell experiment was used to observe the invasion ability of the cancer cells before and after the transfection, and the parent and the transfected cells were injected into nude mice to observe the tumor growth.
Results: After the stable transfection with Her2 shRNA, mRNA and protein levels of Her2 gene in SKOV3 and SKOV3.ip1 cells were remarkably reduced. The expression of mRNA were (68.0 +/- 3.1)%, (40.8 +/- 2.0)%, (99.9 +/- 1.3)%, (42.4 +/- 2.5)%. The expression of protein were (72.1 +/- 3.4)%, (36.4 +/- 1.5)%, (98.2 +/- 1.7)%, (40.7 +/- 2.1)%. The invasion ability into basilar membrane of the transfected cells was greatly reduced compared with the parent cells. The invasion cell numbers were 7.6 +/- 1.1, 1.8 +/- 0.8, 36.2 +/- 9.7, 15.7 +/- 7.2. The growth rate of the planted tumors was lower in transfected groups than that of the parent groups.
Conclusions: (1) The expression of Her2 gene in SKOV3 and SKOV3.ip1 cells was remarkably reduced by RNA interference targeting Her2. (2) The biological characters of SKOV3 and SKOV3.ip1 cells are changed when the expression of Her2 gene is reduced by RNA interference.
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