Effect and mechanism of berberine on proliferation and invasion of colon cancer cells
10.7683/xxyxyxb.2024.11.001
- VernacularTitle:小檗碱对结肠癌细胞增殖和侵袭的影响及机制
- Author:
Shu ZHANG
1
;
Shanshan MENG
;
Yunhe ZHANG
;
Jing CAI
Author Information
1. 河南大学第一附属医院病理科,河南 开封 475001
- Keywords:
berberine;
colon cancer;
hypoxia-inducible factor-1α;
cell proliferation;
cell invasion
- From:
Journal of Xinxiang Medical College
2024;41(11):1001-1007
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of berberine on the proliferation and invasion of human colon cancer cells and its mechanism.Methods The paraffin-embedded tumor tissues and intestinal incisal tissues(control tissues)of 96 patients who underwent radical resection of colon cancer at the First Affiliated Hospital of Henan University from January 2021 to December 2022 were selected as the research subjects.The expression of hypoxia-inducible factor-1α(HIF-1α)in tumor tissues and control tissues was detected by using the immunohistochemical method,and the relationship between HIF-1α and clinicopathological features was analyzed.The human colon cancer cell line HCT116 was cultivated in normoxic and hypoxic conditions,respectively;after adhesion,the cells were divided into normoxic group,hypoxic group,normoxic combined with berberine group,and hypoxic combined with berberine group.The cells in the normoxic combined with berberine group and the hypoxic combined with berberine group were treated with 25,50,75,and 100 μmol·L-1 berberine,respectively;while the cells in the normoxic group and the hypoxic group were not treated with any drugs.At 24,48,72,and 96 hours of cultivation,the proliferation of cells in each group was detected by using the thiazolyl blue tetrazolium bromide method,respectively.Based on the above results,the most suitable drug concentration was determined as 75 μmol·L-1.At 24 hours of cultivation,the invasion of cells in the normoxic group,normoxic combined with 75 μmol·L-1 berberine group,hypoxic group,and hypoxic combined with 75 μmol·L-1 berberine group was detected by using the Transwell method;at 48 hours of cultivation,the relative expression levels of HIF-1α and vascular endothelial growth factor(VEGF)proteins in the cells in each group were detected by using Western blot.Results HIF-1α was mainly expressed in the nucleus of colon cancer tissues and control tissues.The positive expression rates of HIF-1α in control tissues and colon cancer tissues were 19.8％(19/96)and 67.7％(65/96),respectively;the positive expression rate of HIF-1α in colon cancer tissues was significantly higher than that in control tissues(x2=11.298,P＜0.05).The expression of HIF-1α in colon cancer tissues was correlated with tumor size,lymph node metastasis,and TNM staging(P＜0.05),but had no correlation with patient gender,age,tumor infiltration depth,and differentiation degree(P＞0.05).The cells in the normoxic group showed logarithmic growth after 24 hours of culture;and the addition of berberine at a concentration of 25 μmol·L-1 had no significant effect on cell proliferation(P＞0.05);compared with 0 μmol·L-1berberine,the addition of berberine at concentrations of 50 μmol·L-1 and 75 μmol·L-1 significantly reduced cell proliferation activity after 48 hours of culture(P＜0.05),and the addition of berberine at a concentration of 100 μmol·L-1 significantly reduced cell proliferation activity after 24 hours of culture(P＜0.05).After 48 hours of culture with berberine at a concentration of 25 μmol·L-1,the proliferation activity of cells in the hypoxic group was significantly higher than that in the 0 μmol·L-1 berberine(P＜0.05);compared with 0 μmol·L-1 berberine,the addition of berberine at concentrations of 50,75,and 100 μmol·L-1 significantly reduced cell proliferation activity after 24 hours of culture(P＜0.05),and the higher the concentration of berberine,the more pronounced the decrease in cell proliferation activity(P＜0.05).At the same concentration of berberine,compared with the normoxic group,the hypoxic group showed a more significant decrease in cell proliferation activity and a more pronounced slowing-down trend in cell proliferation(P＜0.05).There was no statistically significant difference in the number of cells crossing the membrane between the normoxic group and the normoxic combined with 75 μmol·L-1 berberine group(t=0.955,P＞0.05);the number of cells crossing the membrane in the hypoxic combined with 75 μmol·L-1 berberine group was significantly less than that in the hypoxic group(t=5.354,P＜0.05);the number of cells crossing the membrane in the hypoxic group was significantly more than that in the normoxic group(t=2.625,P＜0.05).There was no statistically significant difference in the relative expression levels of HIF-1α and VEGF proteins between the normoxic group and the normoxic combined with 75 μmol·L-1 berberine group(P＞0.05);the relative expression levels of HIF-1α and VEGF proteins in the hypoxic group were significantly higher than those in the normoxic group(P＜0.05);the relative expression levels of HIF-1α and VEGF proteins in the hypoxic combined with 75 μmol·L-1 berberine group were significantly lower than those in the hypoxic group(P＜0.05);there was no statistically significant difference in the relative expression levels of HIF-1α and VEGF proteins between the normoxic combined with 75 μmol·L-1 berberine group and the hypoxic combined with 75 μmol·L-1 berberine group(P＞0.05).Conclusion HIF-1α expression is associated with growth and invasion of colon cancer.Berberine may inhibit the proliferation and invasion of colon cancer cells HCT116 by regulating the expression of HIF-1α and its downstream factor VEGF.
