Combination DLL4 with Jagged1-siRNA can enhance inhibition of the proliferation and invasiveness activity of human gastric carcinoma by Notch1/VEGF pathway.

BACKGROUND/AIMS: To evaluate the effects of adenovirus- mediated gene transfer of DLL4 and Jagged1 siRNA on proliferation and invasion of SGC7901 cells by Notch/ VEGFR pathway.

METHODOLOGY: Plasmid of DLL4 and Jagged1 siRNA were constructed and transfected into SGC7901 cells. siRNA and endostatin (VEGF inhibitor) were designed as the control group. The mRNA and protein expressions of DLL4 and Jagged1 were respectively detected with RT-PCR and western blotting. In order to find out the changes of proliferation and invasion power of SGC7901 cell lines, we analyzed the data by MTT, Boyden chamber and evaluated apoptosis of cell with flow cytometry. We treated BALB/C nude mice with DLL4 and Jagged1-siRNA, and tumor control rate (%) in nude mice was calculated.

RESULTS: DLL4 and Jagged1 siRNA transfections specifically down-regulated the corresponding mRNA and protein levels in SGC7901 cells. The experiment of permeated artificial basal membrane showed that the invasion power of SGC7901 cell lines were on the decline after treatment of Ad-DLL4- Jagged1-siRNA (12.23±3.12 vs. 78.38±17.38, p<0.05). The values of 490nm wavelength light absorption were different in the five groups. The number of alive cells in the group of DLL4-Jagged1-siRNA was lower than others in the 6th d (0.77±0.01 vs. 3.00±0.11 p<0.05). The apoptosis rate of transfected DLL4 and Jagged1 group with FACS were 18.07%±0.98±1.78 and there were significant differences between treated and control groups (18.07%±0.98 vs. 1.08%±0.23, p<0.01). The tumor transplantation experiment in BALB/C nude mice showed that intratumoral injection of DLL4 and Jagged1 siRNA could inhibit tumor growth.

CONCLUSIONS: DLL4 and Jagged1 siRNA gene therapy mediated by adenovirus may be useful for inhibiting growth and invasion of SGC7901 through a Notch/VEGFR pathway. These results provided a novel therapeutic target in preventing gastric cancer cell invasion and metastasis.