| Cancer is the most important public health problems of the people all over the world,China's cancer data show that in 2015,approximately 4.2916 million new cases,about2.8142 million deaths,and the incidence and mortality of lung cancer are at the top,high recurrence and metastasis is the main cause of death.Antitumor effect of shikonin has extensive,mainly inhibited cell proliferation and inducing tumor cell apoptosis and necrosis,Autophagy,and inhibitory effects on tumor cell migration and invasion.Shikonin found in cancer research with weak inducer of cancer drug resistance,and can circumvent drug transporter and apoptotic defect mediated cancer drug resistance.However,the research of shikonin on the inhibition of non-small cell lung cancer metastasis is not enough.According to reports,TIPE2(TNFAIP8L2)not only the negative immune regulatory molecules,also has the role of the tumor suppressor genes by interacting with RGl/RalGDS and Rac1 molecules.TIPE2 play an important role in the invasion and metastasis of tumor cells,and is closely related to the progression of disease and prognosis of survival in non-small cell lung cancer patients.Rac1 inhibitor(NSC23766)as a specific inhibitor of Rac1,inhibits the invasion and metastasis of gefitinib-resistant non-small-cell lung cancer in clinical studies.However the dose concentration is too large,not widely used.So this article takes shikonin as materials,and try to discuss the possibility of inhibition of non small cell lung cancer A549 cell adhesion,invasion and metastasis through TIPE2-Rac1,and the influence of shikonin and Rac1 inhibitor combined effects on the invasion and metastasis of non small cell lung cancer A549 cells,provides a theoretical basis for the development and application of drugs.The main works carried out in this paper are as follows:1.Shikonin inhibits the metastasis of non-small cell lung cancer A549 cells(1)Cell activity of non-small cell lung cancer A549 cells were tested by MTT method after induction of Shikonin in 12 h,24 h,48 h and 72 h;(2)The adhesion ability of A549 cells was detected by cell-adhesion assays.(3)The migration of A549 cells was detected by wound healing assay.(4)The invasion ability of A549 cells was detected by Transwell.(5)The proteins expression of TIPE2 and Rac1 was analyzed by Western Blot.2.Rac1 inhibitor inhibits metastasis of non-small cell lung cancer A549 cells(1)Cell activity of non-small cell lung cancer A549 cells were tested by MTT method afer induction of Rac1 inhibitor in 12 h and 24 h;(2)The adhesion ability of A549 cells was detected by cell-adhesion assays.(3)The invasion ability of A549 cells was detected by Transwell.(4)The proteins expression of Rac1 was analyzed by Western Blot.3.Rac1 inhibitor and shikonin inhibit the metastasis of non-small cell lung cancer A549cells(1)The adhesion ability of A549 cells was detected by cell-adhesion assays.(2)The invasion ability of A549 cells was detected by Transwell.(3)The proteins expression of TIPE2 and Rac1 was analyzed by Western Blot.The results of this study are as follows:1.The treatment of non-small cell lung cancer A549 cells by shikonin in vitro(1)After induction of shikonin in A549 cells on 12 h,24h,48 h and 72 h,the cells activity were changed.With the increase of shikonin concentration,the A549 cells activity decreased,and showed a dose-and time-dependent manner;(2)Through the cell-adhesion assays,we know that with the increase of shikonin concentration,the adhesion rate of A549 cells decreased.(3)The migration of the cells was detected by scratch test.With the increase of shikonin concentration,the relative migration distance of A549 cells decreased gradually,and showed a dose-and time-dependent manner;(4)Through Transwell assays,we know that with the increase of shikonin concentration,the number of cells into the lower chamber gradually reduced.(5)Through Western Blot Assay,we know that shikonin can significantly increased expression of TIPE2 protein in non-small cell lung cancer A549 cells,and reduced expression of Rac1.2.The treatment of non-small cell lung cancer A549 cells by Rac1 inhibitor in vitro(1)After induction of Rac1 inhibitor in A549 cells on 12 h,and 24 h,the cells activity were changed.With the increase of Rac1 inhibitor concentration,the A549 cells activity decreased,and showed a dose-and time-dependent manner;(2)Through the cell-adhesion assays,we know that with the increase of Rac1 inhibitor concentration,the adhesion rate of A549 cells decreased.(3)Through Transwell assays,we know that with the increase of Rac1 inhibitor concentration,the number of cells into the lower chamber gradually reduced.(4)Through Western Blot Assay,we know that Rac1 inhibitor can significantly increased expression of Rac1 protein in non-small cell lung cancer A549 cells.3.The treatment of non-small cell lung cancer A549 cells by Rac1 inhibitor and Shikonin(1)Through the cell-adhesion assays,we know that Rac1 inhibitor and shikonin combination can significantly reduce the adhesion of non-small cell lung cancer A549 cells,and with the increase of drug concentration,and the adhesion ability decreased gradually;(2)Through Transwell assays,we know that shikonin combined with Rac1 inhibitor can significantly inhibit non-small cell lung cancer A549 cell invasion,and with the increase of drug concentration,and the number of cell invasion decreased gradually;(3)Through Western Blot Assay,we know that Rac1 inhibitors and shikonin combined significantly increased the expression of TIPE2 protein and decrease the expression of Rac1 in non-small cell lung cancer A549 cells.In summary,shikonin inhibits the metastasis of non-small cell lung cancer A549 cells.The inhibition of Rac1 expression can inhibit the migration and invasion of A549 cells by increasing the expression of TIPE2.Rac1 inhibitor and shikonin can significantly increase the expression of TIPE2 protein in non-small cell lung cancer A549 cells,and reduce the expression of Rac1 protein to inhibit the metastasis of A549 cells.Shikonin can increase the sensitivity of Rac1 inhibitors,which lays a theoretical foundation and experimental basis for the clinical treatment of tumor.In this review,shikonin inhibits migration and invasion of non-small cell lung cancer A549 cells.Shikonin can inhibit the metastasis of A549 cells by increasing theexpression of TIPE2 and inhibiting the expression of Rac1.Rac1 inhibitor and shikonin combined significantly increased the expression of TIPE2 protein in non-small cell lung cancer A549 cells,and reduce the expression of Rac1 protein.Shikonin can increase the sensitivity of Rac1 inhibitors,which provides the theoretical foundation and experimental basis for the clinical treatment of tumor. |
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