Inhibitory effect of Notch1 siRNA on proliferation and tumor growth of malignant melanoma cells in vivo and in vitro

Objective To observe the inhibit effeet of siRNA on Notch1 in mouse malignant melanoma cells in vivo and vitro. Method We used siRNA targeting Notch1 gene to transfect melanoma cell line B16F1and induced RNA interference. RT-PCR,Western blotting,MTT method and cell cloning assay in vitro were used to detect the expression of Notch1 gene and protein expression,cell proliferation and cell clone formation changes. Then we transfected or inoculated the control cells subcutaneously into syngeneic C57BL /6 mice,and in the intratumoral process,the corresponding siRNA was injected. The tumor growth was recorded and the growth curve was drawn. After that,we executed the animal to get tumor and compared the tumor volume. Meanwhile,HE and immunohistochemical staining were used to observe the pathological changes of tumor tissue and the expression of Notch1 protein. Results The cell proliferation,clone formation rate and tumor formation in mice were inhibited after Notch1 siRNA transfecting B16F1 cells( P 0. 01),and immunohistochemistry showed that Notch1 protein positive cell percentage of the siNotch1 transfected group was lower than that of control group( all P 0. 01). Conclusions Targeting Notch1 gene siRNA can inhibit the proliferation and growth of mouse melanoma cells by decreasing the expression of Notch1,which can be used as a new method for the treatment of malignant melanoma.