| Objective:Previous proteomic study found that Jinfu'an Tang(JFAT)couldn't only down-regulate the expression of p120ctn in mice lewis lung cancer tissue,but also reduce the levels of phosphorylation on p120ctn in the A549 cell line.And p120ctn also is a key binding partner of transcription factor Kaiso,which is closely related with tunor metastasis,so we could speculate that JFAT might inhibit invasion and metastasis though regulating p120ctn and transcription factor Kaiso-mediated induction in none-small lung cancer.This research would explore the effects of proliferation,invasion and metastasis by JFAT via the vitro experiments in the several human lung cancer cell lines,and we can further study the expression of p120ctn,p120ctn subtypes,its related transcription factor Kaiso and the phosphorylation of serine 288 on p120ctn.So we would explore the mechanism of JFAT through affect the above the signal molecules to inhibit the invasion and metastasis of lung cancer.This study may reveal the molecular mechanisms of invasion and metastasis in tumor cells and provide theoretical and experimental evidence of TCM suppress invasion and metastasis in tumor cells.Methods:Using JFAT tool drugs,human lung adenocarcinoma cell lines A549,H1299,H1650 and human squamous carcinoma cell line H520 as target cells,cell counting kit8 test proliferation of the lung cancer cell lines;Trans well test the invasion and metastasis in the lung cancer cell lines;immunofluorescence observe the location and expression of p120ctn and Kaiso in H1650,H1299 and H520 cells;western blot test expression of p120ctn,p120ctn isform 1A and isform 3A,its related transcription factor Kaiso and the phosphorylation of serine 288 on p120ctn in different kinds of lung cancer cell lines.Results:1.Expriment in vitro showed that JFAT could inhibit the growth and proliferation of human lung adenocarcinoma cell lines A549,H1299,H1650 and human lung squamous carcinoma cell line H520,using cell counting kit8,and this kind of inhibition had a certain time and concentration dependency(p<0.05).And then,in the positive control groups,the same results could be observed in the four different lung cancer cell lines(p<0.05).2.Trans well experiments results revealed that transmenbrane cell numbers of low level,median level and high level JFAT groups were less than the negative control group(p<0.05),and the invasion and metastasis decreased by the concentration dependency.Moreover,the same results could be tested in the positive control group(p<0.05).3.Results of immunofluorescence uncovered in the blank control group,p120ctn in lung squamous carcinoma cell line H520 and lung adenocarcinoma cell lines H1650 was mainly located and significantly high expressed in the cytoplasm and cytomembrane,but pl20ctn in H1299 cell was mainly expressed in the cytoplasm and cell nucleus.We also found that Kaiso blank with control group in H1299 cells,was mainly distributed in cell nucleus,but in H1650 and H520 cell lines was often located in cytoplasm.However,when adding JFAT,p120ctn and Kaiso in H520 and H1299 cell lines significantly decreased(p<0.05),and in cell line H1650,the expression of p120ctn also decreased but the Kaiso up-regulated in JFAT groups.4.Western blot results showed that JFAT could significantly reduced the expression of p120ctn,pl20ctn isform 1A and related transcription factor Kaiso in H1299?A549 and H520 cell lines,and the similar results of p120ctn in the JFAT groups were detected in H1650 cell.Neverthless,we tested the opposite results in JFAT groups,namely Kaiso protein were up-regulated in H1650 cell.And also,the phosphorylation of serine 288 on p120ctn in all cell lines was repressed afer adding JFAT.Conclusion:1.In the vitro research,we found JFAT could inhibit the growth and proliferation of human lung adenocarcinoma cell lines A549,H1299,H1650 and human lung squamous carcinoma cell line H520,and then this kind of inhibition had a certain time and concentration dependency.Furthermore,JFAT also could suppress the invasion and metastasis in lung cancer cells.2.We could speculate that JFAT might inhibit growth,proliferation,invasion and metastasis though regulating p120ctn,p120ctn isform 1A and transcription factor Kaiso-mediated induction in lung cancer cell lines.3.We would also find that the phosphorylation of serine 288 was repressed in lung cancer cells with JFAT intervented and then it could inhibit invasion and metastasis of lung cancer. |
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