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The Effect of DHA Combined with 5-FU on the Proliferation and Apoptosis in Gastric SGC7901 Cells by Rho/ROCK
Wang Keke1, Jiang Yiyan2, Hong Dan1, Lou Zhefeng1, Jin Longjin1*
1School of Laboratory Medicine and Life Science, Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou Medical University, Wenzhou 325035,China; 2The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China
Abstract: The objective of the present study is to explore the synergistic effect of docosahexaenoic acid (DHA) and 5-fluorouracil (5-FU) on the human gastric cancer cell line SGC7901 and its underlying mechanism. The experiments were divided into four groups: control group， DHA group， 5-FU group and combination treatment group. The inhibition of gastric cancer cell proliferation was determined by CCK-8 assay after stimulated 24， 48 and 72 h. Flow cytometric analysis was used to assess cell apoptosis. Real-time PCR and Western blot were used to detect the mRNA and protein levels of RhoA， RhoC and ROCK1 genes， respectively. The results showed 5-FU could increasingly suppress the proliferation of SGC7901 cells with higher doses and longer response time. However， the inhibitory effect， which is not significant with lower doses， appeared significantly in higher doses treated with DHA alone. The inhibitory effect increased when combining 40 μg/mL DHA and 16 μg/mL 5-FU. Compared with the control group， the pro-apoptotic effect with 40 μg/mL DHA was not obvious， 60 μg/mL DHA mainly increased the late apoptosis cells and 16 μg/mL 5-FU induced early apoptosis effect respectively. Co-treatment with DHA and 5-FU enhanced the post-apoptotic effects. The levels of RhoA mRNA decreased in DHA group and 5-FU group while the levels of RhoC mRNA increased in 5-FU group and combination treatment group compared with control group. Meanwhile， the levels of RhoA and RhoC proteins declined with DHA treatment and 5-FU decreased the levels of RhoA and ROCK1 proteins. Combination treatment group resulted in a significantly decrease of RhoA protein level. These results showed that DHA and 5-FU synergetic inhibited the proliferation of gastric SGC7901 cells and affected the cell apoptosis in different periods. Mechanism of synergistically affecting cell proliferation and apoptosis might work through reducing the level of RhoA protein in SGC7901 cells.
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