The Molecular Mechanism Of Baicalin Inhibiting Colorectal Cancer Through NCAPD2 And NCAPD3

Background:Colorectal ancer is one of the most common gastrointestinal tumors.Although surgical resection of primary or metastatic tumors is the best treatment for patients,most colorectal cancer patients cannot achieve complete radical surgery.Therefore,radiotherapy and cchemotherapy have important significance in the treatment of colorectal cancer.Cell mitosis is the basis of eukaryotic growth.Non-smc condensingl complex subunit D2 and non-smc condensing ? complex subunit D3 play key roles in in the mitotic process of eukaryotic cells.Our preliminary experiments showed that baicalin can inhibit the expression of NCAPD2/NCAPD3,but the molecular mechanism of baicalin inhibiting colorectal cancer through NCAPD2/NCAPD3 has not been clarified,which needs further exploration.Aim:1.To observe the expression of NCAPD2 and NCAPD3 in human colorectal cancer tissues,and to clarify the relationship between NCAPD2 and NCAPD3 in the occurrence and development of colorectal cancer.2.To explore the molecular mechanism of baicalin inhibiting colorectal cancer by NCAPD2 and NCAPD3 through the intervention of baicalin in vitro culture of different human colorectal cancer cells.Methods:1.Frozen samples from normal colorectal tissues and tumor tissues were selected to extract tissue proteins,and the expression levels of NCAPD2/NCAPD3 in normal tissues and tumor tissues were detected by Western blot.2.The expression of NCAPD2/NCAPD3 genes in different tissues was detected by immunohistochemistry using wax samples from normal colorectal tissues,ulcerative colitis tissues and colorectal tumor tissues.3.Cell culture and proliferation.Human colon cancer cells HT-29 at 37 ? and 5%C02 saturated humidity cell cultivation box.When the cells were grown to cover the bottom of the bottle,the cells were passaged in a ratio of 1:5 and continued to be cultured.Human colorectal cancer cell line(ht-29)was used as the experimental material to observe the effects of baicalin of different concentrations on proliferation,differentiation and apoptosis of ht-29 cells4.Statistical treatment,SPSS21.0 statistical software was used,and all indexes were expressed as M SD.T-test was used to compare the mean of the two samples,and one-way analysis of variance was used for the comparison between the groups.Results:1.The expression of NCAPD2 and NCAPD3 in colorectal cancer tissues was higher than that in normal colorectal tissues by Western blot(P<0.01)2.Immunohistochemical detection showed that the gene expression of'NCAPD2/NCAPD3 in ulcerative colitis tissues was significantly higher than that in normal tissues(P<0.05),and the expression of NCAPD2/NCAPD3 in colorectal cancer tissues was higher than that in normal tissues,and the difference was statistically significant(P<0.05).3.The addition of baicalin in concentrations of 0.2,0.3,0.4,0.5,0.6 and 0.8 mg/mL to ht-29 cells significantly inhibited the survival rate of ht-29 cells after 48 hours and 72 hours(P<0.05).In the part of cell differentiation,there was a tendency to increase cell differentiation with increasing concentration.Flow cytometry analysis of cell cycle changes showed that adding baicalin with concentrations of 0.15,0.3 and 0.6 mg/mL to cells for 48 hours could increase the percentage of cell number in sub-g0 stage(P<0.05).After 36 hours,adding 0.3 and 0.6 mg/mL baicalin to the cells could induce apoptosis,but at 0.6 mg/mL,it also increased apoptosis.The protein expression of NCAPD2 and NCAPD3 after the intervention of different concentrations of baicalin solution for 48h culture,as well as the protein expression after the addition of inhibitors,confirmed that baicalin may inhibit the growth of tumor cells ht-29 and promote the apoptosis and necrosis of tumor cells by inhibiting the expression of NCAPD2 and NCAPD3.Conclusion:1.NCAPD2 and NCAPD3 are highly expressed in ulcerative colitis and colorectal cancer tissues,and NCAPD2 and NCAPD3 are related to the occurrence and development of colorectal cancer2.Baicalin can inhibit human colorectal cancer cell lines(ht-29)through NCAPD2 and NCAPD3,which can inhibit cell growth and induce apoptosis.