MiRNA-211 triggers an autophagy-dependent apoptosis in cervical cancer cells: regulation of Bcl-2.

Cervical cancer is a significant cause of morbidity and mortality in gynecological malignancies. Although autophagy plays a critical role in affecting cell apoptosis and proliferation, the role of hsa-miR-211-5p (miR-211) in modulating autophagy of cervical cancer cells remains unclear. In the current study, the level of miR-211 was downregulated in cervical cancer specimens, compared to the paired para-carcinoma tissues. While Bcl-2 was upregulated, LC3-II/I was decreased in the tumors, indicating inhibited apoptosis and autophagy. The forced expression of miR-211 inhibited proliferation, and promoted apoptosis in SiHa cervical cancer cells, evidenced by increased expression of apoptotic proteins, caspase-3, and PARP. While the miR-211 inhibitor exerted reverse effects on C-33A cervical cancer cells. Further, miR-211 induced autophagy in cervical cancer cells, as manifested by the presence of LC3 puncta, increased LC3-II/I and Beclin1 levels, and decreased p62 level. The miR-211-induced apoptosis was alleviated by an autophagy inhibitor 3-methyladenine (3-MA). In addition, Bcl-2 was identified as a target of miR-211. Besides, the apoptosis and autophagy triggered by miR-211 were attenuated by Bcl-2 in SiHa cells. In summary, our work indicates that miR-211 induced autophagy and autophagy-dependent apoptosis by regulating Bcl-2 in cervical cancer cells, which provided further understanding of autophagy in cervical carcinogenesis.

Effect of autophagy on the resveratrol-induced apoptosis of ovarian cancer SKOV3 cells.

OBJECTIVE: This study aims to evaluate the relationship between apoptosis and autophagy induced by resveratrol (Res) in SKOV3 human ovarian cancer cell lines. METHODS: The experiment was divided into four groups: normal control group, Res group, Res combined with autophagy inhibitor 3-methyladenine (Res+3-MA) group, and Z-VAD-FMK group. SKOV3 cells were cultured and treated with Res and 3-MA for 24 hours. The processing concentration of Res was screened out by the Cell Counting Kit-8 (CCK8) assay. The cell survival rate was measured by the CCK8 assay. The expression of bule-associated protein light chain 3 beta 2 (LC3-II) and Beclin-1 was detected using Western blot analysis. The percentages of apoptotic and autophagic cells were analyzed using flow cytometry. RESULTS: The cell survival rate significantly decreased as Res concentration increased, and the differences were statistically significant (P < 0.05). The processing concentration of Res was 25 µmol/L. After treatment with Res for 24 hours, the expression levels of autophagy-related protein LC3 and Beclin-l were significantly higher than in the other groups. Furthermore, the expression of LC3 and Beclin-l significantly declined in the Res+3-MA group compared with the Res group. However, the percentage of autophagic cells significantly decreased from 37.0% ± 4.24% to 6.1% ± 0.28%, and the percentage of apoptotic cells significantly increased from 24% ± 4.55% to 67.0% ± 4.3%; and the differences were statistically significant ( P < 0.05). CONCLUSION: Res can induce autophagy to inhibit apoptosis in tumor SKOV3 cells, and inhibition of Res+3-MA could not only enhance the effects of chemotherapy but also prevent normal cells from tumorigenesis.

Ovarian cancer variant rs2072590 is associated with HOXD1 and HOXD3 gene expression.

Ovarian cancer (OC) is a common cancer in women and the leading cause of deaths from gynaecological malignancies in the world. In addition to the candidate gene approach to identify OC susceptibility genes, the genome-wide association study (GWAS) methods have reported new variants that are associated with OC risk. The minor allele of rs2072590 at 2q31 was associated with an increased OC risk, and was primarily significant for serous subtype. The OC risk-associated SNP rs2072590 lies in non-coding DNA downstream of HOXD3 and upstream of HOXD1, and it tags SNPs in the HOXD3 3' UTR. We think that the non-coding rs2072590 variant may contribute to OC susceptibility by regulating the gene expression of HOXD1 and HOXD3. In order to investigate this association, we performed a bioinformatics analysis by a functional annotation of rs2072590 variant using RegulomeDB (version 1.1), HaploReg (version 4.1), and PhenoScanner (version 1.1). Using HaploReg, we identified 19 genetic variants tagged by rs2072590 variant with with r2 >= 0.8. Using RegulomeDB, we identified that three genetic variants are likely to affect TF binding + any motif + DNase Footprint + DNase peak. Other genetic variants are likely to affect TF binding + DNase peak. Using PhenoScanner (version 1.1), we identified that these 19 genetic variants could significantly regulate the expression of nearby genes, especially the HOXD1 and HOXD3 in human ovary tissue.

Association between higher expression of interleukin-8 (IL-8) and haplotype -353A/-251A/+678T of IL-8 gene with preeclampsia: A case-control study.

Preeclampsia (PE) is a common pregnancy-specific disorder associated with significant maternal and fetal morbidity and mortality worldwide.The present study was performed to investigate the role of a CXC chemokine interleukin-8 (IL-8), in the pathogenesis of PE. IL-8 expression levels were assessed in placental and serum samples from 160 pregnant women with PE (N = 68 severe, 92 mild) and 140 healthy donors.Results from enzyme-linked immunosorbent assay showed that the concentration of serum IL-8 in PE patients (180.27 ±â€Š5.81 ng/L) was significantly higher than that in healthy controls (41.57 ±â€Š5.67 ng/L). Patients with severe PE had even higher serum IL-8 levels. Similar messenger RNA and protein expression patterns of IL-8 in placental tissues were confirmed by quantitative real-time polymerase chain reaction and immunohistochemical assay (N = 30 each in the mild PE, severe PE, and control groups). In addition, single nucleotide polymorphisms of IL-8 gene were detected with polymerase chain reaction-restricted fragment length polymorphism/SSP. The frequency of IL-8-251A allele was significantly higher than that in controls (58.4% vs 48.9%, P < 0.05). The occurrence frequency of haplotype -353A/-251A/+678T (AAT) in PE subjects was 27.2% as compared to 21.9% in the control participants (P < 0.05).Our study reveals that IL-8 expression is positively associated with the severity of PE. Presence of haplotype AAT in pregnant women appears to be a risk factor for PE.

Associations between antioxidant vitamins and the risk of invasive cervical cancer in Chinese women: A case-control study.

Previous studies on the associations between dietary antioxidant vitamins and the risk of cervical cancer remain inconsistent, and little evidence is available for serum antioxidant vitamins, which provide more accurate measurements of these nutrients. We conducted a case-control study of 458 incident cases with invasive cervical cancer and 742 controls to assess the effects of diet or serum antioxidant vitamins. Higher serum antioxidant vitamins were associated with a lower risk of cervical cancer after adjusting for potential confounders. The odds ratios (ORs) for the highest (vs. lowest) quartile were 0.66 (95% confidence interval [CI] = 0.46-0.93; P = 0.024) for α-carotene, 0.63 (95% CI = 0.45-0.90; P = 0.006) for ß-carotene, 0.53 (95% CI = 0.37-0.74; P < 0.001) for vitamin E, and 0.48 (95% CI = 0.33-0.69; P < 0.001) for vitamin C. Dietary intakes of vitamins E and C were inversely associated with the risk of cervical cancer. Risk of cervical cancer from serum antioxidant vitamins was more evident in passive smokers than non-passive smokers. These findings indicated that antioxidant vitamins (mainly α-carotene, ß-carotene, and vitamins E and C) might be beneficial in reducing the risk of invasive cervical cancer in Chinese women, especially in passive smokers.

RNAi-mediated knockdown of the CLN3 gene inhibits proliferation and promotes apoptosis in drug-resistant ovarian cancer cells.

CLN3 is a recently identified anti-apoptotic gene, which has been demonstrated to be highly expressed in a diverse range of cancer cell lines, including ovarian cancer. In the present study, RNA interference, mediated by a lentivirus expressing CLN3 short hairpin RNA (shRNA) was utilized to knockdown the expression of CLN3 in the A2780 human ovarian cancer cell line, and its cisplatin­resistant and carboplatin­resistant sublines, A2780/DDP and A2780/CBP cells. It was revealed that the mRNA and protein expression levels of CLN3 were significantly reduced in the CLN3­specific shRNA­transduced cells, compared with the untransduced and control shRNA­transduced cells. In addition, specific knockdown of CLN3 in these cells inhibited cell proliferation and led to cell cycle arrest at the G0/G1 phase, with eventual apoptosis. CLN3 knockdown caused increases in the levels of Bax, FAX, cleaved­caspase 3, cleaved­caspase 8 and cleaved­RARP, but decreased the level of Bcl­2. Finally, it was observed that CLN3 depletion markedly reduced the half maximum inhibitory concentration in the A2780/DDP and A2780/CBP cells. Taken together, these data suggested that CLN3 is involved in tumorigenesis and drug resistance in ovarian cancer, and may serve as a promising therapeutic target for its treatment.

Nuclear export of VP19C is not essential for replication of herpes simplex virus type 1.

BACKGROUND: Herpes simplex virus (HSV) type 1 has a 152 kb double-stranded DNA genome that may encode more than 80 gene products, many of which remain uncharacterized. The HSV-1 triplex is a complex of three protein subunits, VP19C and a dimer of VP23 that is essential for capsid assembly. Previous studies have demonstrated that HSV-1 VP19C contains an atypical nuclear localization signal and a functional nuclear export signal (NES), which are both important for the nucleocytoplasmic shuttling of VP19C. However, whether the VP19C NES is required for efficient HSV-1 production is unknown. FINDINGS: In the present study, a VP19C NES-mutated recombinant virus was generated by using bacterial artificial chromosome recombineering technology to investigate the role of VP19C nuclear export in HSV-1 replication. Our results demonstrate that the growth curves, plaque areas, subcellular localization and viral gene expression are indistinguishable between the VP19C NES-mutated virus and the wild-type virus. CONCLUSIONS: Our findings reported herein indicate abrogation of the nuclear export of VP19C did not affect HSV-1 replication and viral gene expression.

Association of homocysteine, asymmetric dimethylarginine, and nitric oxide with preeclampsia.

PURPOSE: Endothelial dysfunction underlies the pathogenesis of preeclampsia, but its mechanism has not yet been completely understood. In this study we have aimed to measure homocysteine (Hcy), asymmetric dimethylarginine (ADMA), and nitric oxide (NO) levels as endothelial dysfunction markers in preeclamptic women. METHODS: Control-case study with 62 preeclamptic patients and 30 controls without pregnancy complications was conducted. Plasma total Hcy, determined by capillary column gas chromatography/mass spectrometry (GC/MS), was correlated with serum ADMA (determined by liquid chromatography/tandem mass spectrometry using (13)C(6)-L: -arginine as the internal standard) and NO (analyzed by GC/MS). RESULTS: There was a highly significant increase in the plasma concentration of homocysteine (P < 0.001) and ADMA (P < 0.001) and a highly significant decrease in the plasma concentration of nitric oxide (P < 0.001) among the preeclamptic patients. The differences were more significant between mild and severe preeclampsia, with and without eclampsia, with and without HELLP (hemolysis, elevated serum level of liver enzymes, and low platelets). In the combined patients and control groups a highly significant positive correlation was found between the plasma concentrations of homocysteine and ADMA (r = 0.853, P < 0.001). In addition, significant negative correlations were detected between the plasma concentrations of nitric oxide and the plasma concentration of homocysteine (r = -0.870, P < 0.001) and ADMA (r = -0.895, P < 0.001). These significant correlations were found to persist, even when they were restricted to the preeclamptic patients. CONCLUSIONS: The homocysteine-ADMA-NO may be at least partly responsible for etiology in preeclampsia and could be regarded as markers for the severity of the disease. Therefore, L: -arginine may represent a novel therapy for the treatment of preeclampsia.

Genetic polymorphisms of interleukin 8 and risk of ulcerative colitis in the Chinese population.

BACKGROUND: Interleukin-8 (IL-8), a CXC chemokine that recruits and activates inflammatory cells, plays a critical role in the pathogenesis of ulcerative colitis (UC). There are no studies on the association of single nucleotide polymorphisms (SNPs) of the IL-8 gene with the risk of UC. METHODS: All 162 unrelated UC patients and 203 control subjects were analyzed for 5 IL-8 SNPs ((-845 (T/C), -738 (T/A), -353 (A/T), -251 (T/A) and +678 (T/C)) using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) assay and PCR-sequence-specific primers (SSP) method. Serum IL-8 concentrations were measured in all subjects. RESULTS: Individual SNPs were not associated with risk for UC. However, the frequency of -353A/-251A/+678T haplotype was significantly higher in UC patients than in healthy controls (OR=1.454, p=0.036). By subgroup analyses, this haplotype tended to be more common in severe UC patients than in those with mild-to-moderate disease (OR=2.281, p=0.027). Furthermore, patients with AAT diplotype showed significantly increased serum IL-8 concentrations than those with other diplotypes (p<0.001). CONCLUSION: These results suggest that IL-8 is a novel susceptibility gene to UC in Chinese UC patients, and furthermore, that IL-8 polymorphisms may be related to severe clinical subtype of UC.

Polymorphisms of the macrophage inflammatory protein 1 alpha and ApoE genes are associated with ulcerative colitis.

BACKGROUND AND AIMS: An increased production of macrophage inflammatory proteins 1 alpha (MIP-1alpha) has been reported to be associated with ulcerative colitis (UC). We investigated whether a polymorphism site in MIP-1alpha was associated with UC in a Chinese population. Additionally, considering the abnormal lipoprotein metabolism in subjects with UC, we also sought to determine whether genetic variation in the apolipoprotein E (ApoE) gene may play a role in the development of UC. MATERIALS AND METHODS: We examined the MIP-1alpha -906 (TA)(4)/(TA)(6) polymorphism and the ApoE polymorphism in a cohort of 162 unrelated UC patients and 220 healthy controls by using restriction fragment length polymorphism assay. RESULTS: A significantly increased frequency of the MIP-1alpha -906 (TA)(6)/(TA)(6) genotype (P = 0.0031, odds ratio [OR] = 1.851, 95% confidence interval [CI] 1.228-2.791), as well as of the ApoE epsilon4+ genotype (P < 0.001, OR = 2.869, 95% CI 1.768-4.657), in patients with UC was proven. Moreover, the carriage of both MIP-1alpha -906 (TA)(6)/(TA)(6) genotype and ApoE epsilon4+ genotype confers greater risk for the development of UC (P < 0.001, OR = 5.432, 95% CI 2.761-10.689). CONCLUSION: These findings suggest that variation in the MIP-1alpha and ApoE genes and their interaction may increase susceptibility to UC. Identifying these novel susceptibility genes, as well as their interactions, will help our understanding of the disease mechanisms of UC and may identify targets for developing novel treatment measures.

[Study of teratogenicity of hyperglycemia on neural tube defects and antagonistic effect of taurine].

OBJECTIVE: To explore the molecular mechanism of neural tube defects (NTDs) caused by hyperglycemia and thiadiazole and the antagonistic effect of taurine. METHODS: The pregnant mice were divided into hyperglycemia groups, thiadiazole group, taurine groups and control groups. The mRNA and the protein of Pax3 or Cx43 gene were detected respectively by reverse transcription-polymerase chain reaction assay and immunohistochemical method. RESULTS: As compared with mice treated by thiadiazole-stomach-perfusing, NTDs were significantly increased from mice treated with glucose-injection when blood glucose levels were >or= 13.4 mmol/L. Elevated glucose and thiadiazole could cause changes in Pax3 and Cx43 expression. Hyperglycemia had stronger developmental toxicity on mice embryos. Expression of Pax3 (mRNA 0.97 +/- 0.20, protein 0.11 +/- 0.02) in hyperglycemia group was significantly decreased, while expression of Cx43 (mRNA 7.05 +/- 1.63, protein 0.94 +/- 0.05) was significantly increased, and the relationship of dose-effect was demonstrated. In the thiadiazole group, the expression of Cx43 (mRNA 6.96 +/- 0.73, protein 0.92 +/- 0.12) was significantly stronger than control groups, but there were no significant differences in expression of Pax3 between thiadiazole and its control groups. Both of their teratogenicity could be antagonized by taurine. CONCLUSIONS: This study suggests that congenital malformation associated with diabetic pregnancy is caused by disruption of regulatory genes, Pax3 and Cx43 expression in embryo in response to elevated glucose. Thiadiazole can only disturb the regulation of Cx43 gene causing NTDs. Taurine can correct the disruption caused by the two teratogens.

[Expression of Pax3 and Cx43 in NTD embryos induced by hyperglycemia].

OBJECTIVE: To find out the molecular mechanisms of togenesis, especially neural tube defects (NTDs) caused by hyperglycemia and thiadiazole. METHODS: Mice were raised with a ratio of 2:1 between females and males. Forty-five pregnant female mice were randomly divided into 5 groups: blank control group (without any treatment), high-glucose group (treated with subcutaneous injection of 25% glucose), glucose-control group (n = 12, treated with subcutaneous injection of 5% glucose), thiadiazole group (treated with gastric perfusion of thiadiazole), and thiadiazole-control group (n = 11, treated with the solvent of thiadiazole). The blood sugar was examined among the mice of the first three groups on the 8th gestational day. All of the mice were killed on the 15th gestational day and the embryos were taken. The existence of teras was observed. RT-PCR was used to detect the expression of Pax3 and of Cx43 genes and immunohistochemistry was used to examine the Pax3 and Cx43 proteins in the embryos. RESULTS: The blood sugar of the pregnant mice in the high-glucose group was 13.4 mmol/L +/- 0.8 mmol/L, significantly higher than those in the 2 control groups (4.9 mmol/L +/- 0.4 mmol/L and 4.8 mmol/L +/- 0.4 mmol/L respectively, both P < 0.01). The teras rate, absorbed embryo rate and still embryo rate in high-glucose group were significantly higher than those in the 2 control groups (all P < 0.01). The embryo weight of the high-glucose group was significantly lower. The expression of Pax3 in the hyperglycemia group was significantly lower than those in the 2 control groups (both P < 0.01), while the expression of Cx43 in high-glucose group was significantly higher than those in the 2 control groups (both P < 0.01). In the thiadiazole group, the expression of Pax3 was not significantly different from, and the expression of Cx43 was significantly higher than those in the 2 control groups. CONCLUSION: Hyperglycemia is induced in nondiabetic pregnant mice by subcutaneous glucose injection, and elevated glucose appears to be critical in the diabetic embryopathy. Hyperglycemic episodes disturb the essential embryonic control genes: decrease the expression of Pax3 gene and increase the expression of Cx43 gene, thus causing congenital defects. Thiadiazole induces NTDs by increasing the expression of Cx43.