| Objective To investigate the effects of different concentrations of zedoary turmeric oil on the proliferation of SW620 human colon cancer cells and liver metastasis and OPN gene expression.To confirm that zedoary turmeric oil can inhibit the proliferation of colon cancer cells,and by down-regulating the expression of OPN gene to reduce the liver metastasis from colon cancer.Methods In vivo experiment:Establishment of liver metastasis model of colon cancer in mice by removing spleen.18 male BALB/c nude mice were selected for splenectomy.0.1 mL SW620 cell suspension was injected under the splenic capsule during operation.After operation,they were randomly divided into three groups:high concentration group(200μL/mL),low concentration group(100μL/mL)and control group(5%glucose solution),6 in each group.The nude mice in the experimental group were given 200 μL/mice every other day.The control group was given the same amount of 5%glucose solution.After 15 consecutive doses,killed the mice.The liver tissue was taken and divided into two parts.One part was stored in 10%formalin and the other in liquid nitrogen.Through HE staining was used to observe the pathological changes of liver metastasis of colon cancer in each group.The expression of OPN mRNA and protein was detected by RT-PCR and Western-blot.Results In vitro experiment:Compared with negative control group,OD value of 12.5 uL/mL zedoary turmeric oil group had no significant difference(P>0.05),there was no significant difference.Compared with the negative control group,the OD values of the other experimental groups were significantly smaller(P<0.01),and the difference was statistically significant The OD values of 400 uL/mL zedoary turmeric oil group and 200 uL/mL zedoary turmeric oil group were significantly smaller than those of 100 uL/mL zedoary turmeric oil group(P<0.01),the difference is statistically significant.In vivo experiment:Liver metastasis model of colon cancer in mice was established by splenectomy.After killed,tumors were scattered in the liver of 18 nude mice,and the number of tumors in the experimental group was significantly less than that in the control group.HE staining of pathological sections of the specimens showed that they were colon cancer cells,suggesting that the model was successful.HE staining further indicated thatexperimental group could significantly inhibit the proliferation of colon cancer SW620 cells and reduce the occurrence of liver metastasis.Detection of OPN mRNA expression in each group by RT-PCR that the expression of OPN in low dose group was significantly lower than that in negative control group(p<0.05),and the expression of OPN in high dose group was further lower than that in low dose group(p<0.05).Western-blot assay showed that the expression of OPN in experimental group was lower than that in control group.Compared with the control group,the expression of OPN in the low dose group(100μL/mL)was lower than that in the control group(p>0.05),with no significant difference.Compared with the control group,the expression of OPN in the high dose group(200μL/mL)was significantly decreased(p<0.05),and the difference was statistically significant.Compared with the low dose group(100μL/mL),the expression of OPN in the high dose group(200pμL/mL)was lower than that in the low dose group(p>0.05).,but the difference was no statistically significant.Conclusion:1.Zedoary turmeric oil can significantly inhibit the proliferation of human colon cancer SW620 cells.Within a certain concentration,the inhibitory effect was positively correlated with the concentration.When the concentration of zedoary turmeric oil was 200 μL/mL,the inhibitory effect on the proliferation of human colon cancer SW620 cells was the most obvious.2.Zedoary turmeric oil can reduce the occurrence of liver metastasis of colon cancer by down-regulating the expression of OPN and OPN mRNA in metastatic foci. |
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