Inhibitory Effect and Mechanism of Sishenwan-containing Serum on Aerobic Glycolysis in Human Colon Cancer Cells
10.13422/j.cnki.syfjx.20230130
- VernacularTitle:四神丸含药血清抑制人结肠癌细胞有氧糖酵解的效应及机制
- Author:
Yifang JIANG
1
;
Ya HUANG
1
;
Chong XIAO
1
;
Shuwen ZHOU
1
;
Lili ZHENG
1
;
Fengming YOU
1
Author Information
1. Traditional Chinese Medicine（TCM） Regulating Metabolic Diseases Key Laboratory of Sichuan Province， Hospital of Chengdu University of TCM， Chengdu 610075， China
- Publication Type:Journal Article
- Keywords:
Sishenwan;
colon cancer;
glycolysis;
methyltransferase-like 3 （MettL3）;
N6-methyladenosine （m6A） RNA methylation
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2023;29(19):26-33
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effect and mechanism of Sishenwan-containing serum on aerobic glycolysis in human colon cancer HCT116 cells. MethodCell counting kit-8 （CCK-8） was used to detect the cell viability of colon cancer HCT116 cells after treatment with Sishenwan-containing serum （2.5%， 5%， and 10%） for 24， 48， 72 h. The concentration of lactic acid， the content of intracellular glucose， and the activity of hexokinase （HK） and fructose-6-phosphate kinase （PFK） in the cell culture medium were detected by the micro-method. The content of glucose transporter 1 （GluT1） mRNA was detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression of GluT1 and methyltransferase-like 3 （MettL3） was detected by Western blot. The expression of GluT1 in cells was detected by immunofluorescence and the level of N6-methyladenosine （m6A） RNA methylation was detected by colorimetry. ResultCompared with the normal serum， 2.5%， 5%， and 10% Sishenwan-containing serum had no significant effect on the viability of HCT116 cells at 24 h， while 10% Sishenwan-containing serum showed a significant inhibitory effect on the viability of HCT116 cells at 48 h （P<0.05）. Hence， 10% Sishenwan-containing serum was used in subsequent experiments， and the intervention time was 48 h. Compared with the normal serum， 10% Sishenwan-containing serum could reduce lactate production （P<0.05）， down-regulate glucose uptake （P<0.05）， and blunt the activities of HK and PFK， the key rate-limiting enzymes of glycolysis （P<0.05）. Meanwhile， 10% Sishenwan-containing serum could decrease the expression of GluT1 protein （P<0.01） and mRNA （P<0.05） and reduce the proportion of cells expressing GluT1 （P<0.01）. Compared with the normal serum， Sishenwan-containing serum also decreased the protein content of MettL3 （P<0.05） and the methylation level of m6A RNA （P<0.01）. ConclusionSishenwan can inhibit glycolysis in colon cancer cells， and its inhibitory mechanism may be related to reducing MettL3 overexpression， inhibiting m6A RNA methylation， and down-regulating GluT1 and the activities of intracellular aerobic glycolysis-related enzymes such as HK and PFK.
