Network pharmacology and molecular docking reveal the mechanism of resveratrol in oral squamous cell carcinoma treatment / 口腔疾病防治

Objective@#To explore the molecular mechanism of resveratrol (RES) in the treatment of oral squamous cell carcinoma (OSCC) through the use of biological information methods such as network pharmacology and molecular docking and to provide a theoretical reference for the clinical application of RES in the treatment of OSCC.@*Methods@#The Swiss Target Prediction(http://www.swisstargetprediction.ch), SEA (http://sea.bkslab.org)database, and Pharm mapper database(http://lilab-ecust.cn) were used to retrieve RES-related targets, and the DISGENET (www.disgenet.org), OMIM (https://omim.org) and GeneCards (https://www.genecards.org) databases were used to screen OSCC disease targets. The intersection of drugs and disease targets was determined, and Cytoscape 3.7.2 software was used to construct a "drug-diseasetarget pathway" network. The Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database was used to construct a target protein interaction network, and the DAVID database was used for enrichment analysis of key proteins. Finally, molecular docking validation of key proteins was performed using AutoDock and PyMOL. The enrichment analysis and molecular docking results were integrated to predict the possible molecular mechanisms of RES treatment in OSCC; western blot was used to determine the effect of resveratrol at different concentrations (50, 100) μmol/L on the expression of Src tyrosine kinase (SRC), epidermal growth factor receptor (EGFR), estrogen receptor gene 1 (ESR1), and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) signaling pathway proteins in OSCC HSC-3 cells.@*Results@#A total of 243 targets of RES drugs and 6 094 targets of OSCC were identified. A total of 116 potential common targets were obtained by intersecting drugs with disease targets. These potential targets mainly participate in biological processes such as in vivo protein self-phosphorylation, peptide tyrosine phosphorylation, transmembrane receptor protein tyrosine kinase signaling pathway, and positive regulation of RNA polymerase Ⅱ promoter transcription, and they interfere with the PI3K/AKT signaling pathway to exert anti-OSCC effects. The docking results of resveratrol with OSCC molecules indicated that key targets, such as EGFR, ESR1, and SRC, have good binding activity. The results of cell-based experiments showed that resveratrol inhibited the protein expression of SRC, EGFR, ESR1, p-PI3K, and p-AKT in HSC-3 cells in a dose-dependent manner.@*Conclusion@#RES can inhibit the expression of its targets EGFR, ESR1, SRC, p-PI3K, and p-AKT in OSCC cells.

Association between estrogen receptor gene polymorphism and lipid metabolism in the Ningxia Hui population / 西安交通大学学报(医学版)

Objective To study the distribution of rs2234693 and rs9340799 of estrogen receptor (ER)gene and the relationship between them and lipid metabolism in the Ningxia Hui group,China.Methods We used cluster sampling method to select 5 3 3 cases of Ningxia Yinchuan communities of Hui. SNP genotyping was performed using the Sequenom MassARRAY platform.Results ① The frequencies of the distribution of each genotype of rs2234693 in Ningxia Hui population were the same as those in Chinese Han and Dai,Americans, Europeans,and Japanese (P>0.05).The distribution of each genotype of rs9340799,especially AA genotype,was different from that in Europeans (P0.05).Conclusion ER genes distribution in the Hui nationality of Ningxia is significantly different from that in other races;rs2234693 has no obvious relationship with the occurrence of dyslipidemia while GG genotype in SNP rs9340799 significantly increases the risk of lipid metabolism disorders.

Estrogen receptor ERα36 gene silencing impacts the expression of growth-associated protein in PC12 cells / 中华行为医学与脑科学杂志

Objective To investigate the effects of ERα36 (a novel subtype of estrogen receptor alpha) on growth and proliferation in PC12 cells via examining the expression of growth-associated protein in differentiated PC12 cells after ERα36 gene silencing.Methods Transfection of ERα36-shRNA plasmid into PC12 cells was performed to establish the ERoα36 gene silencing cells model (PC12-36L1,PC12-36L2).Immunocytofluorescence was used to examine the expression of ERα36,and Western blot was used to analyze the expression of PCNA,cyclinD1 and MAPK in the PC12 cells.Results ① ERα36 was expressed in both cell types(PC12-36C1,PC12-36L1 and PC12-36L2).Compared with PC12-36C1,PC12-36L2 cells(OD value were respectively 0.95±0.05,0.78±0.10),PC12-36L1 cells significantly decreased expression of ERα36(OD value 0.47±0.12,P＜0.01).② Compared with PC12 and PC12-36C1 cells,PC12-36L1 cells were significantly higher expression of PCNA,CyclinD1 and p-MAPK(P＜0.01)(OD value of PCNA,CyclinD1 and p-MAPK:PC12 cells were respectively 1.00±0.05,1.00± ±0.11,1.00±0.05,PC12-36C1 cells were respectively 1.09±0.15,0.92±0.23,1.12± 0.08,PC12-36L1 cells were respectively 1.74±0.12,2.20±0.25,1.77±0.06).Conclusion ERα36 gene silencing can promote the growth and proliferation in PC12 cells.It suggests that the lower expression of ERα36 may be related to the diseases in nervous system such as brain tumor.

Prevalence of ERα-397 PvuII C/T, ERα-351 XbaI A/G and PGR PROGINS polymorphisms in Brazilian breast cancer-unaffected women

Polymorphisms of hormone receptor genes have been linked to modifications in reproductive factors and to an increased risk of breast cancer (BC). In the present study, we have determined the allelic and genotypic frequencies of the ERα-397 PvuII C/T, ERα-351 XbaI A/G and PGR PROGINS polymorphisms and investigated their relationship with mammographic density, body mass index (BMI) and other risk factors for BC. A consecutive and unselected sample of 750 Brazilian BC-unaffected women enrolled in a mammography screening program was recruited. The distribution of PGR PROGINS genotypic frequencies was 72.5, 25.5 and 2.0% for A1A1, A1A2 and A2A2, respectively, which was equivalent to that encountered in other studies with healthy women. The distribution of ERα genotypes was: ERα-397 PvuII C/T: 32.3% TT, 47.5% TC, and 20.2% CC; ERα-351 XbaI A/G: 46.3% AA, 41.7% AG and 12.0% GG. ERα haplotypes were 53.5% PX, 14.3% Px, 0.3% pX, and 32.0% px. These were significantly different from most previously published reports worldwide (P < 0.05). Overall, the PGR PROGINS genotypes A2A2 and A1A2 were associated with fatty and moderately fatty breast tissue. The same genotypes were also associated with a high BMI in postmenopausal women. In addition, the ERα-351 XbaI GG genotype was associated with menarche ≥12 years (P = 0.02). ERα and PGR polymorphisms have a phenotypic effect and may play an important role in BC risk determination. Finally, if confirmed in BC patients, these associations could have important implications for mammographic screening and strategies and may be helpful to identify women at higher risk for the disease.

Estrogen Receptor Dinucleotide Polymorphism in Patients with Endometriosis / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the estrogen receptor dinucleotide repeat polymorphism with the risk of endometriosis. DESIGN: Case-control study. METHODS: One hundred fifty-one women with surgically or histologically diagnosed endometriosis of stages I-IV (ASRM, 1997) were recruited, and 137 patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. Dinucleotide repeat polymorphism of the estrogen receptor gene was assessed by fluorescent PCR with gene scan analysis. Allele frequencies of dinucleotide repeat polymorphism of the estrogen receptor gene were evaluated. RESULTS: Fifteen alleles of the estrogen receptor dinucleotide repeat polymorphism were found in subjects: from 12 repeats to 27 repeats except 26 repeats. There was no statistically significant difference in the allele distribution of dinucleotide repeat polymorphism between patients with endometriosis and controls. However, patients with stage I or II endometriosis (n=51) showed a higher incidence of alleles with fewer (TA)n repeats (12-15 repeats) compared with controls (67.6% vs 52.9%, p=0.010, odds ratio=1.860). CONCLUSION: These results suggest that dinucleotide repeat polymorphism of the estrogen receptor gene is associated with the risk of minimal or mild endometriosis in the Korean population.

Effects of Estrogen Receptor Polymorphisms on Bone Markers and Serum Lipid Levels / 대한진단검사의학회지

BACKGROUND: In post-menopausal women, osteoporosis and cardiovascular diseases which are partly due to estrogen deficiency, occur more common than in pre-menopause women. Estrogen action is supposed to be mediated by an estrogen receptor (ER) and two polymorphisms of the ER gene in particular, Pvu II and Xba I, have been described for several years for genetic association studies. Authors have investigated the frequencies and patterns of the ER gene polymorphisms and their association with bone markers and lipid levels. METHODS: For 121 women who visited the health promotion center of Kyungpook National University Hospital, the ER gene polymorphisms were determined by the Pvu II and Xba I restriction enzymes following polymerase chain reaction. RESULTS: The distributions of ER Pvu II and Xba I restriction fragment length polymorphisms were as follows: PP 15.7%, Pp 47.9%, pp 36.4% and XX 5.8%, Xx 31.4%, xx 62.8%, respectively. And in a combination of two polymorphisms, ppxx was the most common, followed by PpXx, Ppxx, PPXx, PPXX and PPxx in that order. No significant genotypic differences were found in bone mineral density, bone markers and menopausal status. LDL cholesterol and triglyceride levels were significantly different by genotypes in premenopausal women (P<0.05). CONCLUSIONS: The results suggest that ER polymorphisms might be associated with LDL cholesterol and triglyceride levels. Further evaluation in a larger population would be helpful to determine the effects of ER polymorphisms on lipid metabolism and therapeutic trial for cardiovascular diseases in women.

Estrogen Receptor Gene PvuII and XbaI Polymorphism in Patients with Endometriosis / 대한산부인과학회잡지

OBJECTIVE: To explore the association of the estrogen receptor PvuII and XbaI polymorphism with endometriosis. METHODS: One hundred sixty women with surgically or histologically diagnosed endometriosis of stages I-IV, and 142 patients with no evidence of endometriosis by laparoscopy or laparotomy served as control. Frequency and distribution of PvuII and XbaI polymorphisms for estrogen receptor gene were evaluated. RESULTS: There was no statistically significant difference in the allele distribution of PvuII polymorphism between the patients and the controls (pp of 35%, pP of 51%, PP of 14% vs. 42%, 44%, 15%, p>0.1); or in the frequency of the positive PvuII allele (0.61 vs. 0.63, p>0.1). And no significant difference was also observed in the allele distribution of XbaI polymorphism between the patients and the controls (xx of 66%, xX of 29%, XX of 5% vs. 68%, 30%, 1%, p>0.1); or in the frequency of the positive XbaI allele (0.80 vs. 0.83, p>0.1). CONCLUSION: These results suggest that the PvuII or XbaI polymorphism of the estrogen receptor gene is not associated with the risk for endometriosis in the Korean population.

Association Between Peak Bone Mass and Genetic Polymorphisms of the Vitamin D Receptor, Estrogen Receptor, and Type I Collagen 1 Genes in Healthy Young Korean Women / 대한내분비학회지

BACKGROUND: Genetic suggest that strongest effect is observed in the premenopausal peak bone mass, which become less with age. However, the evaluation of candidate genes polymorphisms has been most frequently done in postmenopausal women and the results have been controversial. Therefore, we studied the possible association of the peak bone mass and candidate for osteoporosis genes polymorphism in premenopausal women. METHODS: The associations between BMD and polymorphisms of the vitamin D receptor (3'-end region by BsmI restriction enzyme and start codon by FokI restriction enzyme), estrogen receptor (by PvuII and XbaI restriction enzyme), and type I collagen 1 (Sp1 binding site by MscI and BalI restriction enzyme) genes were examined in 100 healthy young Korean women who had a peak bone mass (age 20-35 years). Bone mineral densities were measured by dual energy X-ray absorptiometry (DEXA). Dietary calcium intake was also measured using a food frequency questionnaire. RESULTS: The frequencies of the B allele of the vitamin D receptor gene BsmI polymorphism and the X allele in the estrogen receptor gene, XbaI polymorphisms were lower in Koreans than those in Caucasians. The allelic frequencies of the vitamin vitamin D receptor gene FokI polymorphism and the estrogen receptor gene PvuII polymorphism were similar to those of Caucasians. No significant association was found between BMD and the vitamin D receptor genotype according to BsmI or FokI polymorphisms. There was also no significant relation between the PvuII or XbaI polymorphisms of the estrogen receptor gene and BMD. The associations between BMD and cross-genotypes combining the vitamin D receptor gene (BsmI and FokI) and estrogen receptor gene (PvuII and XbaI) polymorphisms were also analyzed. Among the subjects who lacked the Bf haplotype of the vitamin D receptor gene, the BMD of the femoral neck area was significantly higher in subjects lacking Px haplotypes of the estrogen receptor gene than in those having Px haplotype (p < 0.05). When dietary calcium intake was taken into consideration, there were significant differences in BMD according to the cross-genotype in the group having a low calcium intake (< 500 mg/day). The subjects that lacked the Bf and Px haplotypes had a significantly higher BMD in the femoral neck (p < 0.01), Ward's triangle (p < 0.05), and in the trochanteric area (p < 0.05) than those who lacked Bf but a Px haplotype. We did not find a polymorphism in the Sp1 binding site of the type I collagen 1 gene in our subjects. CONCLUSION: These data suggest that a complex interaction of vitamin D and the estrogen receptor gene with the dietary calcium intake, rather than a polymorphism of a single gene, may influence peak bone mass in healthy young Korean women.

Estrogen Receptor Gene Polymorphism, Urinary Estrogen Metabolites and Bone Mineral Density in Korean Postmenopausal Women / 대한내분비학회지

Background: Estrogen status is important for maintaining the homeostasis of bone. Estrogen has direct effects on bone cells, through binding to the high-affinity estrogen receptor. Several recent studies suggest that there might be genetically determined variations in biosynthesis and function of estrogen receptor in postmenopausal osteoporosis. Also the main cause of postmenopausal osteoporosis is decreased level of serum estrogen, whereas there had been some suggestion that the remaining estrogen have some effect on bone metabolism after menopause. We investigated the relationship between estrogen receptor gene PvulI polymorphism and bone mineral density(BMD), and the relationship between 18 urinary metabolites of estrogen and BMD in Korean postmeno- pausal osteoporosis. Methods: We examined the PvuII polymorphism of the estrogen receptor gene in 5' upstream region and the first intron by restrietion frapnent length polymorphism analysis in 62 postmeno- pausal wornen, BMD was measured by DEXA. The urinary estrogen metabolites were determined by GC/MS(Gas Chromatography-Mass Spectrometry) at Korean Institute of Science and Techno- logy Doping Control Center. Results: BMD of the spine and the femoral neck correlated with body weight, height, body mass index as we expected. There was no polymorphism of PvuII restriction site on 5 upstream region of estrogen receptor gene. Whereas the prevalen~ee of the PP, Pp, pp genotype in the first intron of estrogen receptor was 12.9%, 45.2%, 41.9%, respectively. But, there was no correlation between PvuII genotype and the spinel and femoral neck BMD. 2(OH)E2 among 18 urinary metabolites of estrogen, showed a negative correlation with the spinal and femoral neck BMD(r =-0.2551, p revealed a positive correlation with the spinal BMD(r =0.3057, p<0.05). In stepwise multiple regression analysis, body weight, 2(OH)E2, 16a(OH)E1, 2(Meo)E1 were independent predictors of the spinal bone density, and body weight and 2(OH)E2 were independent predictors of the femoral neck bone density. Conclusion: These results suggested that restrietion fragment length polymorphism analysis of the estrogen receptor gene with PvuII restriction enzyme was not helpful for early detection of patients at risk of developing osteoporosis. However, the ratio of 16-hydroxylation to 2-hydroxylation of estrogen metabolism was reduced in postmenopausal women and high catecholestrogen formation might be a greater risk factor for osteoporosis.