| Breast cancer,is the most common malignancy in women.New cancer cases are emerging every year in China,more than 2.8 million deaths every year,and the incidence and mortality rate is rising year by year,the severe situation of the sick family,the country and the community has brought enormous burden.Breast cancer as the highest morbidity and mortality of the type of cancer,because of its high transfer rate,resulting in patients with short survival time,poor prognosis,is a problem to be solved.With the advent of a variety of anticancer drugs,the ensuing drug resistance has become a key factor in the impact of anticancer drugs,the need to develop new drugs.Celastrol is a kind of pure natural compound,it has good anti-inflammatory,swelling,detoxification,inhibit tumor growth and metastasis,and some studies show that its tumor suppressor rate exceeds Taxol.The effect of celastrol on anti-tumor cell metastasis has been studied in many signaling pathways and cell lines,but there is still a lack of research on the inhibition of C-myc transfer by Jak1/Stat3 signaling pathway in breast cancer cell MDA-MB-231.C-myc is one of the most important members of the myc gene family,and it is also a kind of adjustable gene,which can make cell proliferation,immortality function,promote cell division,C-myc gene participates in cell metastasis,and the development of many kinds of tumor.The study reported that 20% of human malignancies were associated with the abnormal expression of C-myc,including non-small cell lung cancer,breast cancer,colon cancer,cervical cancer,myeloid leukemia,melanoma,osteosarcoma,malignant glioma and adult-administered cell tumors.C-myc can promote cell proliferation,promote tumor angiogenesis,tumor cell invasion and migration,and promote tumor cell metastasis.C-myc has become a target of current tumor therapy.In this paper,we investigate the following: MTT assay to detect the cellular activity of md-mb-231 cells after a certain period of time under the action of cel and/or EGF.The adhesion ability of cells was detected by cell adhesion experiment.The scratch test detects the cell's migration ability.Transwell detects the ability of cells to invade.Small tube generation experiment is to detect the ability of small tube generation in vitro.Changes in transcription levels of gene Jak1,Stat3 and C-myc were detected in qPCR.The expression of Stat3,C-myc protein level were detected by Western blot.The results of this study are as follows:Effect of celastrol on cell metastasis of Breast cancer:(1)MTT assay showed that the different concentrations of celastrol in treating mda-mb-231 cells after 24 h could significantly inhibit cell activity and dose-dependent.(2)The results of scratch test showed that celastrol inhibited the migration ability of MDA-MB-231 cells significantly.(3)Transwell experimental results showed that celastrol inhibited the invasion ability of MDA-MB-231 cells significantly.(4)The results of cell adhesion experiments showed that celastrol inhibited MDA-MB-231 cell adhesion.(5)The results of small tube formation experiment showed that celastrol inhibited the formation of MDA-MB-231 cells in vitro.(6)The results of Western Blot showed that celastrol inhibited the expression of Stat3 and C-myc protein.Moreover,celastrol inhibited the effect of EGF on Stat3 protein and C-myc protein.(7)qPCR results showed that celastrol inhibited the effect of EGF on the expression of Jak1,Stat3 and C-myc gene.Through the above experiments,we preliminarily obtained the inhibitory effect of celastrol on the invasion and metastasis of breast cancer cells and inhibition of EGF transfer.By inhibiting the promotion of C-myc and EGF to C-myc,celastrol inhibited the metastasis of breast cancer MDA-MB-231 cells in a dose-tolerant manner.Therefore,the study of the effect of celastrol on the anti-breast cancer and the mechanism of inhibition of EGF promoting metastasis provides the basis for the application of celastrol in anti-tumor field,which lays a foundation for clinical cancer treatment of celastrol. |
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