RG108 induces the apoptosis of endometrial cancer Ishikawa cell lines by inhibiting the expression of DNMT3B and demethylation of HMLH1.

OBJECTIVE: The effects of DNA methyltransferase (DNMT) inhibitor RG108 on the proliferation and apoptosis of endometrial cancer was investigated, and whether its mechanism was related to the inhibition of DNMT3B, thereby affecting the human mutL homolog 1 (hMLH1) methylation status and its expression, was further studied. MATERIALS AND METHODS: Culture of human endometrial cancer Ishikawa cell lines: cells grew adhering to the wall in Roswell Park Memorial Institute-1640 (RPMI-1640) medium (supplemented with 10% fetal bovine serum (FBS) and 2 mM L-glutamic acid). After the cells were treated with RG108, the changes in cell viability were detected via methyl thiazolyl tetrazolium (MTT) assay. The effect of RG108 on cell cycle was detected via flow cytometry, and its effect on cell apoptosis was detected via flow cytometry and TUNEL. Moreover, the methylation status of hMLH1gene in endometrial cancer cells was detected via methylation specific-PCR (MSP), and the changes in DNMT3Band hMLH1 expressions were detected via RT-PCR and Western blotting, respectively. RESULTS: MTT results showed that RG108 inhibited the cell viability in a dose-dependent and time-dependent manner. Flow cytometry revealed that RG108 blocked the cell cycle in G2/M phase and promoted the apoptosis, and TUNEL assay further proved that RG108 promoted the apoptosis. It was found in the detection via MSP that the methylated hMLH1 gene was significantly reduced after 72 h of treatment with RG108. Besides, RT-PCR and Western blotting showed that RG108 inhibited the DNMT3B expression and activated the hMLH1 expression. CONCLUSIONS: The demethylation drug RG108 can significantly inhibit the proliferation of endometrial cancer cells, block the cell cycle in the G2/M phase and induce the cell apoptosis, which is a new candidate drug in the treatment of endometrial cancer. RG108 realizes the hMLH1 demethylation and increases the hMLH1 expression through inhibiting the expression of DNMT3B.

MiR-133b regulates the expression of CTGF in epithelial-mesenchymal transition of ovarian cancer.

OBJECTIVE: To explore the role of miR-133b in ovarian cancer and to preliminarily elucidate the mechanism of miR-133b in epithelial-mesenchymal transition (EMT) of ovarian cancer. PATIENTS AND METHODS: MiR-133b was detected in ovarian cancer specimens, and the relationship of miR-133b with each pathological index and clinical index of ovarian cancer was analyzed. The action targets of miR-133b in ovarian cancer were analyzed systematically and studied deeply via the target validation and cell function validation. Finally, the possible reasons of ovarian cancer metastasis were analyzed through the molecular regulation mechanism in EMT of ovarian cancer. RESULTS: The miR-133b level in ovarian cancer was significantly lower than in normal ovarian tissues and benign ovarian tumors (p<0.05). The level of miR-133b in ovarian cancer was related to differentiated degree and lymphatic metastasis. Dual-luciferase assay indicated that connective tissue growth factor (CTGF) was the target gene regulated by miR-133b. Reverse transcriptase-polymerase chain reaction (RT-PCR) as well as Western blot results proved that the expression level of E-cadherin representing the epithelial cell phenotype was increased, while the expression level of vimentin representing the mesenchymal cell phenotype was decreased. Transwell assay confirmed that the migration and invasion abilities of ovarian cancer cells declined after transfection with miR-133b plasmid. After co-transfection with miR-133b and CTGF overexpression plasmids, RT-PCR and Western blotting proved that the expression level of E-cadherin representing the epithelial cell phenotype was decreased, while the expression level of vimentin representing the mesenchymal cell phenotype was increased; transwell assay confirmed that the cell migration and invasion abilities were increased after co-transfection. CONCLUSIONS: The results of this study showed that miR-133b may serve as a new molecular marker of EMT of ovarian cancer, and act as a molecular marker of differentiated degree and lymphatic metastasis of ovarian cancer.

Thoracolumbar kyphosis is associated with compressive vertebral fracture in postmenopausal women.

The main aim of this retrospective cross-sectional study was to examine the relationship between vertebral compression fracture and thoracolumbar Cobb angles. Fracture prevalence was found to be significantly higher for patients with moderate [odds ratio (OR) = 4.78 (2.88-7.95)] or severe kyphosis [OR = 10.7 (5.11-22.40)] than for patients with mild kyphosis. The relationship between degree of thoracolumbar kyphosis and vertebral compression fracture was analyzed. INTRODUCTION: The hypothesis that vertebral compression fracture in women is related to thoracolumbar kyphosis severity was tested, and a clinically important cutoff degree of sagittal thoracolumbar Cobb angle (TLCobb) was determined. METHODS: Demographic data, clinical data, and quantitative computed tomography (QCT) findings were compiled for 212 postmenopausal women with thoracolumbar fracture (study group) and 150 postmenopausal women with degenerative lumbar disease (control group). Group proportions and characteristics were compared with chi-squared tests and unpaired t tests, respectively. RESULTS: In this retrospective cross-sectional study cohort, 17 patients had T11 fractures, 79 had T12 fractures, 89 had L1 fractures, and 27 had L2 fractures. QCT findings and TLCobb differed between the study and control groups (both p < 0.001). No significant differences were found in body mass index (BMI), disk height, or coronal TLCobb. After adjustment for age, BMI, and QCT findings, fracture prevalence was found to be higher in the thoracolumbar kyphosis study group than in the control group [OR = 6.16, 95% confidence interval (CI) 3.88-9.78]. Sagittal TLCobbs of 7.5-15° and >15° were associated with an increased fracture prevalence, with ORs of 4.78 (2.88-7.95) and 10.7 (5.11-22.40), respectively. CONCLUSION: Vertebral fracture prevalence in postmenopausal women was found to be associated with thoracolumbar kyphosis. A TLCobb sagittal angle >15° should be considered an indicator for vertebral fracture assessment.

Increased human epididymis protein 4 in benign gynecological diseases complicated with chronic renal insufficiency patients.

We examined the serum concentration of human epididymis protein (HE4) in patients with benign gynecological diseases complicated with chronic renal deficiency and its significance in the differential diagnosis of benign and malignant gynecological diseases. Serum HE4 and cancer antigen 125 concentrations were detected by chemiluminescence. Clinically or pathologically confirmed gynecological diseases were grouped and retrospectively analyzed, including 50 cases of gynecological benign diseases, 35 cases of non-mucinous epithelial ovarian carcinoma, 36 cases of endometrial adenocarcinoma, 15 cases of gynecological benign diseases patients complicated with chronic renal deficiency, 15 cases of gynecological diseases without chronic renal deficiency, and 30 normal controls. Serum HE4 values in the ovarian cancer group, endometrial cancer group, gynecological benign diseases with chronic renal deficiency group, and chronic renal deficiency group were significantly increased compared with the benign gynecological diseases and normal control groups, showing a significant difference (P < 0.001). A comparison of 4 groups with high HE4 showed that the HE4 level in the 2 groups with renal deficiency were higher than those in the ovarian cancer and endometrial cancer groups, but the difference was not significant (P > 0.05); there was no significant difference between 2 groups with renal deficiency (P > 0.05). Serum concentration of HE4 was high in patients with chronic renal deficiency, which should be distinguished during differential diagnosis of gynecological benign and malignant tumors in patients with chronic renal deficiency to avoid misdiagnosis.