The Role Of TP63 Protein In The Progression Of Colorectal Cancer And The Intervention Of Cryptotanshinone

Objective:To investigate the effects of TP63 protein expression on the survival cycle of colorectal cancer patients and the proliferation,metastasis and apoptosis of colorectal cancer cells.Based on this,intervention with cryptotanshinone was used to investigate the effects of cryptotanshinone on the proliferation,metastasis and apoptosis of colorectal cancer cells based on STAT3/ΔNp63 pathway.Methods:1 Clinical research: Cancer and paracancer specimens were collected from 19 patients with colorectal cancer undergoing surgery.Western blot was used to detect the expression of p53 family proteins in cancer tissues and adjacent tissues of clinical patients,screen valuable proteins,and follow up the postoperative survival cycle of patients to analyze the correlation between the expression of ΔNp63 protein and the survival cycle.2 Cell experiment: The cells were divided into control group,protein overexpression group and representative knockdown group by transfection of protein expression and knockdown.Clonal colony formation assay was performed to detect the number of clonal colonies formed in each group,and to infer the effect of protein expression on the proliferation of colorectal cancer cells.Transwell assay was used to detect the number of migratory and invasive cells in each group,and to infer the effect of protein expression on colorectal cancer cell metastasis.Apoptotic inducers were used to induce cells,and flow cytometry was used to detect the apoptosis rate of proteins in each group,which was used to speculate the influence of protein expression on apoptosis of colorectal cancer cells.Western blot was used to detect the expressions of apoptotic molecules Bcl-2,Bcl-XL,Mcl-1,P-gp and Bax in each group.To evaluate the effect of protein expression on apoptotic molecular expression in colorectal cancer cells.The intervention of colorectal cancer cells with cryptotanshinone was divided into blank control group and each concentration gradient intervention group.CCK-8 was used to determine the activity of colorectal cancer cells after CTS treatment.Clonal colony formation assay was used to determine the effect of cryptotanshinone on the proliferation of colorectal cancer cells.Transwell assay was used to evaluate the effect of cryptotanshinone on colorectal cancer cell metastasis by detecting migration and invasion after intervention.The expression levels of STAT3/ Δ Np63 pathway and apoptotic molecules Bcl-2,Bcl-XL,Mcl-1,P-gp and Bax in colorectal cancer cells after cryptotanshinone intervention were detected by Western blot.Results:1 Clinical research: Western blot analysis showed that there was no difference in the expression of p53 protein and TAp63 protein between the two tissues(P>0.05),while there was remarkable difference in the expression of ΔNp63 protein between the two tissuess(P<0.05).Divided into high and low expression groups with all patients based on ΔNp63 expression levels.Baseline data was no difference between the two groups(P>0.05).Compared with the two groups,the survival period of patients with ΔNp63 high expression group was significantly shortened(P<0.05).2 Cell experiment: The overexpression and knockdown of ΔNp63 protein in colorectal cancer showed that knockdown of Δ Np63 protein decreased the number of clonal colony formation in colorectal cancer cells,while overexpression of ΔNp63 protein increased the number of clonal colony formation in colorectal cancer cells(P<0.05).Tranwell assay found that knockdown of ΔNp63 reduced the number of migrating and invading colorectal cancer cells,while overexpression of ΔNp63 increased the number of migrating and invading colorectal cancer cells(P<0.05).After apoptosis induction,flow cytometry and Western blot showed that knockdown ΔNp63 protein promoted the expression of apoptotic molecular protein Bax by inhibiting the apoptotic inhibitory molecular proteins Bcl-2,Bcl-XL,Mcl-1 and P-gp,and thus promoted the apoptosis of colorectal cancer(P<0.05).Overexpression of ΔNp63 protein can inhibit the apoptosis of colorectal cancer by promoting the expression of apoptosis-inhibiting molecular proteins Bcl-2,Bcl-XL,Mcl-1 and P-gp,and inhibiting the expression of apoptosis-inhibiting molecular protein Bax(P<0.05).CTS was used to interfere with colorectal cancer cells,and it was found that CTS inhibited the number of clonal colonies formed in colorectal cancer cells(P<0.05).Tranwell assay showed that CTS inhibited the number of migrating and invading colorectal cancer cells(P<0.05).Flow cytometry showed that CTS promoted apoptosis of colorectal cancer cells(P<0.05).Western blot showed that CTS inhibited the phosphorylation of STAT3 protein and the expression of Δ Np63 protein in colorectal cancer(P<0.05).CTS inhibited the expression of Bcl-2,Bcl-XL,Mcl-1 and P-gp,and promoted the expression of Bax(P<0.05).Conclusion:1 Δ Np63 protein is highly expressed in colorectal cancer tissues,and this high expression is inversely proportional to patient survival.2 The overexpression of Δ Np63 protein in colorectal cancer cells promoted the proliferation and metastasis of colorectal cancer cells,and promoted the expression of apoptosis-inhibiting molecules to inhibit the apoptosis of colorectal cancer cells.Knockdowning ΔNp63 expression in colorectal cancer cells inhibited the proliferation and metastasis of colorectal cancer cells,and inhibited the expression of apoptosis-inhibiting molecules,thereby promoting the apoptosis of colorectal cancer cells.3 Cryptotanshinone inhibited the proliferation and metastasis of colorectal cancer through STAT3/ΔNp63 pathway,and promoted the expression of apoptotic molecules to induce the apoptosis of colorectal cancer.