| objective1.To study the effects of Huatan Sanjie decoction on the anti-tumor effect of human gastric cancer cells MKN45 and BCG823 in orthotopic gastric tumors of nude mice.2.To explore the mechanism of Huatan Sanjie decoction may be through miR-506-3p targeting ERK-2 / ETS-1 / MMP9-related signaling pathways and its effect on EMT,resulting in anti-gastric cancer.methods1.Real-time quantitative PCR was utilized to detect the expression level of has-miR-506-3p in gastric cancer cell lines(BCG823,SGC7901,MKN45 and MGC803),and two types of gastric cancer cells with relatively high expression were screened out.2.Human gastric cancer cell lines BCG823 and MKN45 formed subcutaneous tumors and were inoculated near the gastric antrum of the pylorus of nude mice to establish tumor-bearing nude mice.The tumor bearing mice were randomly divided among three groups:three groups of each cell,namely the traditional chinese medicine group(Hutan Sanjie decoction),the western medicine group 5-fluorouraci(5-FU)and the normal saline group.3.During the treatment,the general situation and weight changes were regularly monitor.After 6 weeks,the tumor bearing mice were skilled and the tumors were dissected out,the tumor weight was weighed,the tumor suppression rate was calculated,and the presence of metastasis was observed.Fluorescence quantitative PCR and Western-Blot was used to detect the expression of miRNA-506-3p,ETS-1,ERK-2,MMP9 and E-cad mRNA and protein in the tumor tissues of BCG823 and MKN45 cells.Results1.Compared with the western medicine and saline groups,the traditional chinese medicine group can improve the general condition of the tumor bearing mice’s mental state,diet and activity.2.The body weight of tumor bearing mice in each group was changed after treatment.Depending on the weight of the animals,they were:chinese medicine group>saline group>western medicine group.3.Compared with the saline group,the tumor weight difference between the chinese medicine group and the western medicine group was statistically significant(P<0.01).Compared with the western medicine group,the tumor weight of the traditional chinese medicine group was not statistically significant.The tumor inhibition rates of the MKN45 tissue tumor western medicine group and the chinese medicine group were 54.6% and 41.1%,and the BCG823 tissue tumor western medicine group and the chinese medicine group were 57.1% and 46.7%.4.qPCR:MKN45 tissue(1)miR-506-3p:Compared with the saline group,the chinese medicine group and the western medicine group was upregulated(P<0.001);compared with the western medicine group,the chinese medicine group has no statistical significance.(2)ERK-2:Compared with the saline group,the traditional chinese medicine group and western medicine group decreased(P≤0.001);compared with the western medicine group,the traditional chinese medicine group had no statistical significance.(3)ETS-1:Compared with saline group,the chinese medicine group and western medicine group decreased(P < 0.05);compared with western medicine group,chinese medicine group decreased(P < 0.05).(4)MMP9 is not statistically significant in the traditional chinese medicine group and the western medicine group compared with the saline group;compared with the western medicine group,the chinese medicine group is not statistically significant.(5)E-cad:Compared with saline group,the traditional chinese medicine group and western medicine group increased(P<0.05);compared with western medicine group,the chinese medicine group increased(P<0.001).BCG823 tissue(1)miR-506-3P:Compared with the saline group,the chinese medicine group and the western medicine group was up-regulated(P<0.001);compared with the western medicine group,the chinese medicine group was up-regulated(P<0.001).(2)ERK-2:Compared with saline group,the traditional chinese medicine group and western medicine group decreased(P < 0.001);compared with western medicine group,the traditional chinese medicine group decreased(P<0.05).(3)ETS-1:Compared with saline group,the traditional chinese medicine group and western medicine group decreased(P<0.001);compared with western medicine group,the traditional chinese medicine group decreased(P<0.05).(4)MMP9:Compared with saline group,the chinese medicine group and western medicine group decreased(P<0.001);compared with western medicine group,chinese medicine group decreased(P≤0.001).(5)E-cad:Compared with saline group,the chinese medicine group and western medicine group increased(P <0.001);compared with western medicine group,chinese medicine group increased(P<0.001).5.Western-Blot:MKN45 tissue(1)ERK-2:Compared with the saline group,the chinese medicine group and the western medicine group decreased(P<0.05);compared with the western medicine group,the chinese medicine group decreased(P<0.05).(2)ETS-1:Compared with the saline group,the chinese medicine group and the western medicine group decreased(P<0.05);compared with the western medicine group,the chinese medicine group decreased(P<0.05).(3)MMP9:Compared with the saline group,the chinese medicine group and the western medicine group decreased(P<0.05);compared with the western medicine group,the chinese medicine group decreased(P<0.05).(4)E-cad:Compared with the saline group,the chinese medicine group and the western medicine group increased(P<0.05);compared with the western medicine group,the chinese medicine group increased(P<0.05).In BCG823 tissue MMP9:Compared with the saline group,the chinese medicine group and the western medicine group decreased(P<0.001);compared with the western medicine group,the chinese medicine group decreased(P<0.05).The E-cad,ERK-2 and ETS-1 expression results showed the same trend as the MKN45 tissue.conclusions1.Huatan Sanjie decoction can inhibit the growth of human gastric cancer cells MKN45 and BCG823 orthotopic transplanted tumors in nude mice,which can improve the quality of life of tumor-bearing mice.2.Huatan Sanjie decoction can up-regulate miR-506-3p,down-regulate ERK-2,ETS-1,MMP9,and up-regulate E-cad at the same time,so as to inhibit orthotopic transplanted tumors in nude mice growth. |
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