Expression of long noncoding RNA (linc-VLDLR) in extracellular vesicles and its relation to the development and multidrug resistance of esophageal cancer / 解放军医学杂志

Objective To investigate the relationship between the expression of linc-VLDLR in extracellular vesicles (EVs) and the development and drug resistance of esophageal carcinoma.Methods Fifty percent of inhibitory concentration (ICs0) ofadriamycin (ADM) for Eca109 cells was detected by MTT assay,after the treatment of esophageal squamous cell carcinoma Eca109 cell line with different concentrations of ADM for 24h.The culture medium was treated with 3 concentrations of ADM based on the IC50 for 24h for extracting EVs in Eca109 cells.linc-VLDLR mRNA expression in EVs was detected by qRT-PCR assay.ICs0 of ADM for Eca109 cells intervened by EVs for 48h was detected by MTT assay.Cell cycle was detected by FCM and linc-VLDLR and ABCG2mRNA expressions in Eca109 cells were detected by qRT-PCR after the treatment of the EVs for 48h.Results ICs0 of ADM acting on Eca109 cells for 24h was 0.44 ± 0.02μg/ml,so ADM concentrations of 0.2,0.4,0.8μg/ml were choosed in the following studies.EVs were extracted from the supernatant after the treatment of 0,0.2,0.4,0.8μg/ml ADM for 24h and were labeled as EVs1,2,3 and 4 respectively.LincVLDLR mRNA expression in EVs4 was significantly higher than that in EVs1-3 (P＜0.01).ADM ICs0 for Eca109 cells in EVs4 group was significantly higher than that in other groups after the treatment of EVs1-4 on Eca109 cells for 48h (P＜0.05).Flow cytometry results showed that the proliferation index of Eca109 cells in EVs4 group was significantly higher than that in EVs 1-3 and control groups (P＜0.01).Linc-VLDLR and ABCG2 mRNA expression levels in Eca109 cells of EVs4 group were significantly higher than these of EVs1-3 and control groups (P＜0.05).Conclusions High expression of linc-VLDLR and ABCG2 gene in esophageal cancer cells is involved in the formation of esophageal cancer resistance.EVs released by drug-resistant cells can upregulate the expression of ABCG2 in esophageal cancer cells and regulate the drug resistance of esophageal cancer cells,which is related to the linc-VLDLR gene carried by EVs.

The effect of PLCε1 gene silencing on apoptosis and invasion of esophageal carcinoma Eca109 cells and its mechanism / 肿瘤

Objective: To investigate the effect of phospholipase C epsilon 1 (PLCε1) gene silencing on apoptosis and invasion of esophageal carcinoma Eca109 cells and its possible mechanism. Methods: The recombinant vectors targeting short hairpin RNA (shRNA) of PLCε1 (pGenesil-1-PLCε1-shRNA, pGenesil-1-PLCε1-shRNA2 and pGenesil-1-PLCε1-shRNA3) were transfected into Eca109cells by cationic liposome method and to screen out the expression vector with best interference effect. The apoptosis rate and invasive ability of Eca109 cells 48 h after transfection were determined by flow cytometry (FCM) and Transwell chamber method, respectively. After PLCε1 gene silencing, the expression levels of factorassociated suicide (Fas), Fas ligand (FasL), CD44, matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) mRNAs were detected by semi-quantitative reverse transcriptionpolymerase chain reaction (RT-PCR). Results: The pGenesil-1-PLCε1-shRNA2 had the best interference effect on PLCε1 gene. The apoptosis rate of Eca109 cells 48 h after transfection with PLCε1-shRNA2 was (30.27±5.13)%, which was significantly higher than that of the cells transfected with pGenesil-1-NC-shRNA (NC group) [(22.06±4.47)%, P < 0.05]. The number of Eca109 cells across the polycarbonate membrane of Transwell chamber in the NC group and PLCε1-shRNA2 group 48 h after transfection were 82.00±2.00 and 62.67±3.06, respectively. The number of Eca109 cells across the polycarbonate membrane in PLCε1-shRNA2 group was lower than that of the NC group (P < 0.05). The expression level of Fas mRNA in Eca109 cells of PLCε1-shRNA2 group was significantly up-regulated as compared with that of the NC group (P < 0.05), while the expression levels of FasL, MMP-9 and VEGF mRNAs in PLCε1 - shRNA2 group were significantly down-regulated (all P < 0.05). There was no significant difference in CD44 mRNA expression level between the NC group and PLCε1-shRNA2 group. Conclusion: Silencing PLCε1 gene might promote the apoptosis of Eca109 cells by up-regulating the expression of Fas and down-regulating the expression of FasL, and it can also suppress the invasive ability of Eca109 cells by down-regulating the expressions of MMP-9 and VEGF.

Association of single nucleotide polymorphisms in VEGF gene with the risk of endometriosis and adenomyosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of single nucleotide polymorphisms (SNPs) in VEGF gene with the risk of endometriosis and adenomyosis.</p><p><b>METHODS</b>Genotypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 344 endometriosis patients, 174 adenomyosis patients, 360 frequency-matched control women of endometriosis and 199 frequency-matched control women of adenomyosis.</p><p><b>RESULTS</b>No significant difference was found in allele frequencies and genotype distributions of the -460C/T polymorphism between patients (endometriosis and adenomyosis) and control women (all P value > 0.05). However, there were significant differences in genotype and allele distributions of the VEGF -1154G/A polymorphism between patients (endometriosis and adenomyosis) and control women (all P value < 0.05). The genotype frequencies of the VEGF -1154 AA, GA, and GG in endometriosis patients and control women were 1.7%, 28.8%, 69.5% and 5.8%, 32.8%, 61.4%, respectively; and the A and G allele frequencies in the two groups were 16.1%, 83.9% and 22.2%, 77.8%, respectively. The genotype frequencies of the VEGF -1154 AA, GA, and GG in adenomyosis patients and control women were 2.9%, 23.6%, 73.6% and 7.0%, 34.2%, 58.8%, respectively; and the A and G allele frequencies in the two groups were 14.7%, 85.3% and 24.1%, 75.9% respectively. Compared with GA+ AA genotype, GG genotypes could significantly increase the risk of endometriosis (OR:1.43,95%CI:1.05-1.96) and adenomyosis (OR:1.95,95%CI:1.26-3.03).</p><p><b>CONCLUSION</b>The VEGF -1154G/A polymorphism was associated with susceptibility to endometriosis and adenomyosis, and the GG genotype could significantly increase the risk of developing endometriosis and adenomyosis. However, the VEGF -460C/T polymorphism was not associated with susceptibility to endometriosis and adenomyosis in the population studied.</p>

The association of XRCC2 gene polymorphism with susceptibility to esophageal squamous cell carcinoma and gastric cardiac adenocarcinoma in China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the possible association of single nucleotide polymorphism (SNP) at the 41657C/T position and 4234G/C position of X-ray repair cross-complementing gene 2 (XRCC2) with susceptibility to esophageal squamous cell carcinoma (ESCC) and gastric cardiac adenocarcinoma (GCA) in a population of high incidence region, Ci county and She county of Hebei.</p><p><b>METHODS</b>The genotypes of XRCC2 41657C/T and 4234G/C SNPs were detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis in 330 ESCC patients, 254 GCA patients and 629 healthy controls.</p><p><b>RESULTS</b>The genotype frequency of XRCC2 41657C/T in ESCC patients (67.8%, 26.4% and 5.8%) was significantly different from that in controls (68.8%, 28.8% and 2.4%; chi square was 7.43, P was 0.02). Compared with CC genotype, TT genotype significantly increased the risk of developing ESCC (OR=2.12, 95%CI: 1.03-4.35). The genotype (59.9%, 35.8% and 4.3%) and allelotype distributions ofXRCC2 41657C/T in GCA patients were significantly different from that in controls (chi square was 7.46 and 7.23, P was 0.02 and 0.01). Compared with CC genotype, CT genotype significantly increased the risk of developing GCA (OR=1.38, 95%CI: 1.01-1.89). The genotype and allelotype distributions of the 4234G/C SNPs in ESCC and GCA patients were not significantly different from that in controls (all P values were above 0.05). Compared with GG genotype, the CG and CC genotype of XRCC2 4234G/C did not show significant effect on the risk of developing ESCC and GCA. When the two XRCC2 SNPs were combined analyzed, the haplotype distribution in GCA patients was significantly different from that in controls (chi square was 13.28, P was less than 0.01). Compared with 41657C/4234G haplotype, 41657C/4234C and 41657T/4234G haplotypes significantly increased the risk of developing GCA (OR were 1.44 and 1.55, 95%CI were 1.06-1.95 and 1.18-2.02, respectively).</p><p><b>CONCLUSION</b>In high incidence region of Hebei province, we conclude that XRCC2 41657C/T polymorphism has a potential to be a susceptibility factor for ESCC and GCA while XRCC2 4234G/C polymorphism may not provide a useful marker to predict susceptibility to ESCC and GCA. However, the 41657C/4234C and 41657T/4234G haplotypes might increase the risk of developing GCA.</p>