1.Regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16
Suyun BI ; Li LI ; Song XU ; Mengli ZHANG ; Heng GU ; Zhihai ZHOU ; Xu CHEN
Chinese Journal of Dermatology 2018;51(6):421-424
Objective To evaluate the regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16.Methods Cultured COLO16 cells were divided into 6 groups to be treated with lycopene at different concentrations of 0,5,10,15,20,and 25 μmol/L,respectively,for 24 hours (control group and 5,10,15,20,25 μmol/L lycopene groups),followed by estimation of the cell viability by lactate dehydrogenase (LDH) assay.Lycopene at a safe concentration was selected based on the LDH assay,and used for the determination of expression of signaling receptors,and Western blot analysis was performed to measure the expression of key signaling receptor proteins,including epidermal growth factor receptor (EGFR),glucocorticoid receptor (GR),retinoic acid receptor-alpha (RAR-α),retinoid X receptor-alpha (RXR-α),androgen receptor (AR) and progesterone receptor (PR).Statistical analysis was carried out by one-way analysis of variance (ANOVA),Tukey multiple comparison teat and Brown-Forsythe test with the SPSS software.Results After 24-hour treatment with lycopene at different concentrations,there were significant differences in the rate of cell death among these groups (F =13.116,P < 0.05),and the rate of cell death in the 25 μmol/L lycopene group significantly differed from that in the control group (P < 0.05).Therefor,lycopene at concentrations of 5,10 and 20 μmol/L were selected to treat COLO16 and HaCaT cells as well as human epidermal keratinocyte (HEK) for 24 hours in the following experiment.The treatment with lycopene significantly decreased the phosphorylation level of EGFR (P < 0.05),but significantly increased the expression of GR protein (P < 0.05),and showed no significant effects on the protein expression of RAR-α,RXR-α,AR,and PR in COLO16 cells.After 24-hour treatment with lycopene at concentrations of 5,10 and 20 μmol/L,there were no significant changes in the phosphorylation level of EGFR protein or the expression of GR protein in HaCaT cells and HEK (all P > 0.05) compared with those without lycopene treatment.Conclusion Lycopene can decrease the viability of COLO16 cells,inhibit the activation of EGFR protein,and up-regulate the expression of GR,and these effects may be specific for tumor cells.
2.Relationship between forkhead box P3 gene polymorphisms and susceptibility to gastric cancer
Caijuan SI ; Peifen ZHENG ; Xinxin ZHOU ; Mengli GU ; Feng JI
Chinese Journal of Digestion 2018;38(2):105-109
Objective To explore the relationship between forkhead box P3(Fox P3)gene polymorphism and susceptibility to gastric cancer in Chinese Han population.Methods From January 2014 to December 2016,a total of 312 patients with primary gastric cancer and 548 age and gender matched controls were recruited.Polymorphisms of Fox P3 at rs3761548 and rs2232365 loci were analyzed by polymerase chain reaction-restriction fragment length polymorphism techniques(PCR-RFLP).Odd ratio(OR)and 95% confidence interval(CI)were used to analyze the correlation between different genotype and susceptibility to gastric cancer,clinical pathological characteristics of patients with gastric cancer.Results The frequency of AA,AC,and CC genotype of rs3761548 in gastric cancer group and control group were 22.8%(71/312),44.2%(138/312),33.0%(103/312)and 14.8%(81/548),48.5%(266/548),36.7%(201/548),respectively.AA genotype could increase the risk of gastric cancer(OR=1.711,95% CI 1.150 to 2.545,P=0.008).The frequency of A and G allele of gastric cancer group and control group were 44.9%(280/624),55.1%(344/624)and 39.1%(428/1 096),60.9%(668/1 096), respectively.A allele could increase the risk of gastric cancer(OR=1.270,95% CI 1.041 to 1.550,P=0.018).The frequency of AA,AG,and GG genotype of rs2232365 in gastric cancer group and control group were 19.2%(60/312),46.5%(145/312),34.3%(107/312)and 14.1%(77/548),48.7%(267/548),37.2%(204/548),respectively.In recessive model(AA vs AG+GG),AA genotype could increase the risk of gastric cancer(OR=1.456,95% CI 1.005 to 2.110,P=0.046).A allele of rs3761548 was associated with the depth of invasion in gastric wall(χ2 =12.710,P=0.001)and Helicobacter pylori infection(χ2 = 5.613,P= 0.018).A genotype could increase the risk of lymphatic metastasis(χ2 =4.878,P=0.027).Conclusion Fox P3 gene polymorphism is associated with the susceptibility to gastric cancer in Chinese Han population,and can be molecular-maker of poor prognosis of gastric cancer.