| Colorectal cancer(CRC)is a common malignant tumor of digestive system in China.The resistance of colorectal cancer cells to radiotherapy and chemotherapy is the main reason for the reduction of clinical therapeutic effect,tumor recurrence and metastasis,and death of patients.In recent years,there are many scientific hypotheses and experimental evidences for the formation mechanism of its resistance,but at present,no substantial progress has been made and clinical feasible treatment scheme has been provided.By establishing a chemoradiotherapy resistance model of colorectal cancer in vitro,our team confirmed that forkhead box protein Q1(FoxQ1)plays an important role in the chemoradiotherapy resistance of colorectal cancer cells,and found that the chemoradiotherapy sensitivity of chemoradiotherapy resistance cells in vitro is decreased,which may be related to the interaction between foxq1 and Wnt/?-catenin signaling pathway.Therefore,in order to explore the potential relationship between foxq1 and chemoradiotherapy resistance in colorectal cancer,our group plans to use cell transfection technology to inhibit and over express the expression level of foxq1 in chemoradiotherapy resistance cell line of colorectal cancer.QRT-PCR and Western blot experiments respectively analyze whether the expression of ?-Catenin is related to the expression of FoxQ1 at mRNA and protein levels,and analyze the relationship between them.On this basis,this study further explored whether artemether(ART)can reverse the resistance of cancer cells to radiotherapy and chemotherapy through this pathway,in order to provide new clues for the mechanism of resistance of colorectal cancer to radiotherapy and chemotherapy,and also provide solid theoretical and experimental evidence for ART to reverse the resistance of colorectal cancer cells to radiotherapy and chemotherapy at the same time.[Objective]1.To establish a CRT resistant cell model of colorectal cancer HCT116/HT-29 and verify its CRT resistance2.To explore the interaction between FoxQ1,a key gene of CRT resistance,and Wnt/?-Catenin signaling pathway;3.It was found that ART could regulate Wnt/?-Catenin signaling pathway through FoxQ1,and reverse the CRT resistance of CRC cells.[Method]1.Human CRC cell line HT29/HCT116 was treated with 10?mol/L 5-FU and 4Gy 6MV X-ray in vitro;2.MTT cell proliferation experiment,plate cloning experiment,Transwell migration and invasion experiment were used to observe the difference of proliferation,cloning,migration and invasion ability between HT29CRR/HCT116CRR cell line and HT29/HCT116 cell line,analyze the resistance of HT29CRR/HCT116CRR cell line to radiotherapy and chemotherapy at the same time,and identify whether HT29CRR/HCT116CRR cell line was successfully established;3.The expression of foxq1 in HT29CRR and HCT116CRR cell lines was inhibited or over expressed by cell transfection.Each cell line was divided into control group and experimental group,and qRT-PCR and Western blot were used The relationship between FoxQ1 and Wnt/?-catenin signaling pathway was analyzed.It was confirmed that foxq1 has a regulatory effect on Wnt/?-Catenin signaling pathway in CRC chemoradiotherapy resistant cell line.4.MTT assay was used to calculate IC50 of ART on HT29CRR/HCT116CRR cell line FoxQ1 and ?-Catenin of ART treated and untreated CRT resistant colorectal cancer cells HT29CRR/HCT116CRR were detected by qRT-PCR and Western blot To verify whether ART can regulate Wnt/?-catenin signaling pathway through FoxQ1;5.The experimental data were analyzed by SPSS 19.0 software package and expressed by mean±standard deviation(±s).The data between groups were analyzed by one way ANOVA and t test.[Result]1.After 12 concurrent chemoradiotherapy in vitro,successfully established HT29 residual cancer cell line after concurrent chemoradiotherapy;2.MTT cell proliferation experiments,plate cloning experiments,Transwell migration and invasion experiments confirmed that HT29 residual cancer cell lines have concomitant chemoradiotherapy resistance after concurrent chemoradiotherapy,and are concomitant chemoradiotherapy resistant cell lines3.At the mRNA and protein levels,inhibition of FoxQ1 in HT29CRR/HCT116CRR caused a decrease in ?-Catenin expression;overexpression of FoxQ1 caused an increase in ?-Catenin expression,which was statistically significant.4.At the mRNA and protein levels,the expression levels of FoxQ1 and ?-Catenin in the HT29CRR/HCT116CRR artemether treatment group were lower than those in the untreated group and were statistically significant;[Conclusion]1.Application of high-dose shock therapy in vitro can improve the chemoresistance of human colorectal cancer cell line HT29/HCT116 in vitro2.FoxQ1 has radiochemotherapy resistance regulation effect on Wnt/?-Catenin signaling pathway.3.Artemether can regulate Wnt/?-Catenin signaling pathway through foxq1 to reverse the chemoradiotherapy resistance of colorectal cancer. |
PDF Full Text Request