| Objective: Esophageal cancer is one of the malignant diseases that give a serious threat to human life,mainly squamous cell carcinoma.It belongs to the category of Yege in Chinese traditional medicine. Eesophageal cancer is rapid progression,easily metastasis and poor prognosis once pathogenesis,lost opportunity to operation.As radiation and chemotherapy side effects,so it is a great significance to seek traditional Chinese medicine treatment which is safe,effective and non-toxic side effects.The Runzaojiangqi Fang was a modified prescription that had a role in the treatment process of esophageal precancerous lesion from studies.This study focused on the signaling pathways associated protein closely related with tumor cell invasion,took focal adhesion kinase(FAK)and Paxillin as research subjects,observed the Combination Runzaojiangqi Fang pathological changes to esophageal precancerous lesion and expression of cell adhesion-related proteins,provided a theoretical basis on the Combination Runzaojiangqi Fang in the course of preventive treatment of esophageal cancer.Methods: We took C57 BL / 6 mice as research subjects,built esophageal precancerous lesion model induced by condensation of 4-nitroquinoline 1-oxide(4NQO).The mouse were divided into normal control group,model group,positive control group,Chinese medicine treatment group and Chinese medicine prevention group to conduct the impact of the Combination Runzaojiangqi Fang to esophageal precancerous lesion.By HE staining,we observed the pathological changes in different disposal groups.By immunohistochemical method to detect the protein expression situation of Paxillin and FAK,we used RT-PCR method to detect the expression changes of Paxillin and FAK in different disposal groups;then projected the expression by gel imaging system and software,used SPSS17.0 to test the difference among the different treatment groups(P <0.05).1 Physiological conditionsThe mouse in normal group performed lively and normally;In other group the mouse were normal in 2 weeks;some mouse started depression and neck hair removal after two weeks;In the end of the experiment,the mouse began fluffy fur,arched curling and loss of appetite;the mouse in prevention group had a better status.2 HE stainingThe mouse exhibited regular morphology of esophageal tissue in normal group;After induced by 4NQO the esophageal tissue would exhibit seriously hyperplasia in squamous cells;In positive,treatment and prevention group,hyperplasia became attenuated.3 Expression of Paxillin and FAKBy Immunohistochemistry, Paxillin mainly exsisted in the cytoplasm or cell membrane and presented brownish yellow. The expression of Paxillin in the normal group was low.The expression of Paxillin in other groups was different.The expression of Paxillin in the normal group was 0.143 ± 0.007. The expression of Paxillin in the model group was 0.218 ± 0.021,and was significantly high expression than in normal group(P<0.05).The expression in the positive and treatment group were 0.195 ± 0.016 and 0.153 ± 0.015, significantly lower than model group,respectively(P<0.05).The expression in treatment group was lower than the positive group,but there was no significant difference(P>0.05).By Immunohistochemistry,FAK mainly exsisted in the cytoplasm and presented brownish yellow. The expression of FAK in the normal group was low.The expression of FAK in other groups was different.The expression of FAK in the normal group was 0.156 ± 0.013.The expression of FAK in the model group was 0.239 ± 0.031,and was significantly high expression than in nomal group(P<0.05).The expression in the positive and treatment group were 0.185 ± 0.02 and 0.165 ± 0.009, significantly lower than model group,respectively(P<0.05).The expression in treatment group was lower than the positive group,but here was no significant difference between the treatment group and the positive control group(P>0.05).4 Gene expression of Paxillin and FAKA specific band occurred at 334 bp with Paxinllin primers amplified by RT-PCR;The brightest among the amplified bands was the model group and the normal group was the darkest.The positive group,prevention group, treatment group compared with the model group,the amplified bands darker;at the same time, the positive group,treatment group,prevention group was similar in brightness.The expression of Paxillin presented by the ratio of absorbance between paxillin and GAPDH.The results showed:in the model group,the ratio of FAKm RNA/GAPDHm RNA increased significantly compared with the normal group(P<0.05),the positive control group, prevention group,the treatment group was significantly lower than those in the model group(P<0.05),treatment group was significantly higher than those in the positive control group(P<0.05),prevention group and positive control group had no significant difference(P>0.05),prevention group ratio was lower than the treatment group(P<0.05).A specific band occurred at 740 bp with Paxinllin primers amplified by RT-PCR; The brightest among the amplified bands was the model group and the normal group was the darkest.The positive group,prevention group, treatment group compared with the model group,the amplified bands darker;at the same time,the positive group was brighter and prevention group was darker than the treatment group in brightness.There were significantly correlation in the following group:all the other groups compared with the model group;the treatment group was significantly lower than the positive control group(P<0.05),The prevention and the positive group were no significantly correlation(P>0.05),the prevention group had no difference compared with the treatment group(P>0.05).Conclusions:1 4NQO could effectively induce esophageal cell proliferation. Runzaojiangqi Fang could effectively reduced esophageal cell proliferation and had a equivalent effect with ARTA in the reversal of esophageal cancer. Chinese medicine prevention can reduce esophageal precancerous lesion pathologic damage and has a certain advantage.2 Runzaojiangqi Fang could reduce the protein and gene expression of Paxillin and FAK in the esophageal tissue.As a test results of the RT-PCR,it was found that prevention drug had an obvious advantage in inhibiting the gene expression of FAK.ARTA could reverse the esophageal cancer as a effective treatment.The test results by RT-PCR showed that prophylaxis had obvious advantages in gene expression.3 Lower the gene expression of Paxillin and FAK,might be one of effective way of Runzaojiangqi Fang on the suppression of esophageal precancerous lesion progress.A further study is needed about the specific mechanism of Runzaojiangqi Fang. |
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