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Background Arsenic is a toxicant that can affect the expressions of the cellular anti-apoptotic gene BCL-2 and its protein, but the effects of arsenic on BCL-2α and BCL-2\begin{document}$\beta $\end{document} transcripts have not been reported. Objective To investigate the potential effects of arsenic and its metabolites, methylarsonic acid (MMA) and dimethylarsonic acid (DMA), on BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T (total of α and \begin{document}$\beta $\end{document} transcripts) in human bronchial epithelial cells (16HBE) and human lung adenocarcinoma cells (A549). Methods 16HBE cells and A549 cells were randomly divided into three categories of exposure after in vitro culture: single-selected arsenic compound exposure groups with isoconcentration (16HBE cells were treated with 4.5 μmol·L−1 of MMA, DMA, and sodium arsenite, respectively, while A549 cells were treated with 60 μmol·L−1 of MMA, DMA, and sodium arsenite, respectively), sodium arsenite exposure groups with different concentrations (16HBE cells were treated with 1.5, 3.0, and 4.5 μmol·L−1 of sodium arsenite respectively, while A549 cells were treated with 20, 40, and 60 μmol·L−1 of sodium arsenite respectively), and combined exposure groups (i.e. MMA+sodium arsenite, and DMA+sodium arsenite; the exposure concentrations of 16HBE cells were both 1.5 μmol·L−1 and both 4.5 μmol·L−1 respectively, and those of A549 cells were both 20 μmol·L−1 and both 60 μmol·L−1 respectively). Meanwhile, a blank control group was also set up in each exposure category. After 48 h of continuous exposure, the relative expressions of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T in both cells were detected by real-time PCR. Results Regarding the single-selected arsenic compound exposure, in 16HBE cells, the expression levels of BCL-2α and BCL-2T under 4.5 μmol·L−1 MMA treatment were lower than those in their control groups (q=3.27, 2.93, both P<0.05), and the expression levels of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T under 4.5 μmol·L−1 sodium arsenite were lower than those in their respective control groups (q=11.06, 3.65, 10.70, all P<0.05). In A549 cells, the expression level of BCL-2T treated with 60 μmol·L−1 DMA was lower than that in the control group (q=3.12, P<0.05), and the expression levels of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T treated with 60 μmol·L−1 sodium arsenite were lower than those in their respective control groups (q=7.59, 7.27, 8.06, all P<0.05). Regarding the sodium arsenite exposure: 16HBE cells treated with 1.5 μmol·L−1 sodium arsenite had a lower expression level of BCL-2α and a higher expression level of BCL-2\begin{document}$\beta $\end{document} than those in their respective control groups (q=6.06, 11.92, both P<0.05); the expression level of BCL-2α under 3.0 μmol·L−1 sodium arsenite was lower than that in the control group (q=12.72, P<0.05); and under 4.5 μmol·L−1 sodium arsenite treatment, the expression levels of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T were lower than those in their respective control groups (q=15.72, 6.79, 6.62, all P<0.05). The expression levels of BCL-2α gradually decreased with increasing concentrations of sodium arsenite (Fα trend=144.80, P<0.001), while BCL-2\begin{document}$\beta $\end{document} and BCL-2T decreased in a dose-dependent manner in the range of 1.5-4.5 μmol·L−1 (F\begin{document}${}_{\beta } $\end{document} trend=135.40, FT trend=38.24, both P<0.001). In A549 cells, the expression levels of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T under each concentration of sodium arsenite treatments were lower than those in their respective control groups (all P<0.05); the results of further trend tests showed that their expression levels gradually decreased with increasing concentrations of sodium arsenite (Fα trend =31.97, F\begin{document}${}_{\beta} $\end{document} trend=549.50, FT trend=252.40, all P<0.001). Regarding the combined exposure, under MMA+sodium arsenite treatment at both 60 μmol·L−1, the expression levels of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T in A549 cells were higher than those in their respective control groups (q=6.37, 14.91, 5.33, all P<0.05); under DMA+sodium arsenite treatment at both 60 μmol·L−1, their expression levels in A549 cells were also higher than those in their respective control group (q=8.60, 17.29, 6.91, all P<0.05). Conclusion Exposure to a high concentration (16HBE: 4.5 μmol·L−1, A549: 60 μmol·L−1) of a single arsenic metabolite has no effect on BCL-2 transcripts in 16HBE cells and A549 cells. Exposure to a low concentration (1.5 μmol·L−1) of sodium arsenite alone would decrease the expression level of BCL-2α and increase the expression level of BCL-2\begin{document}$\beta $\end{document} in 16HBE cells, and exposure to all designed concentrations of sodium arsenite alone would decrease the expressions of all transcripts in A549 cells. The combined exposure to high concentrations (both 60 μmol·L−1) of MMA plus sodium arsenite or high concentrations (both 60 μmol·L−1) of DMA plus sodium arsenite would increase the expressions of BCL-2α, BCL-2\begin{document}$\beta $\end{document}, and BCL-2T in A549 cells, which are different from the effects presented by single exposure.
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Objective:To investigate the correlation of mRNA expression levels and DNA methylation levels of Alu-mediated p21 transcriptional regulator (APTR) with hepatitis B virus infection.Methods:One hundred patients with HBV infection admitted in Affiliated Hospital of Yunnan University during January to December 2019 were enrolled in the study, including 50 patients with chronic hepatitis B (CHB group) and 50 asymptomatic HBV carriers (ASC group); and 50 healthy subjects were also enrolled as the healthy control group. The DNA methylation levels of APTR gene were detected by methylation-sensitive high-resolution melting (MS-HRM); the expression levels of APTR mRNA were detected by fluorescence real-time quantitative PCR (qRT-PCR). Pearson correlation or Spearman rank correlation was used for correlation analysis.Results:There were significant differences in the APTR DNA methylation levels among the CHB, ASC and healthy control groups {[12.02 (9.30, 23.32)]%, [10.02 (8.46, 17.44)]% and [8.86 (7.82, 11.57)]%, χ2=13.360, P<0.01}. The APTR DNA methylation levels were significantly higher in CHB group than those in healthy control group( Z=31.480, P<0.01). There were significant differences in the APTR mRNA expression levels among CHB, ASC and healthy control groups (2.38±1.41, 5.78±2.78 and 5.70±2.66, F=33.720, P<0.01). The APTR mRNA expression levels were significantly lower in CHB group than those in healthy control and ASC groups ( t=7.808 and 7.724, both P<0.01). Correlation analysis showed that the DNA methylation level of APTR gene was negatively correlated with mRNA expression levels ( r=-0.305, P<0.01) in all subjects. The DNA methylation level of APTR gene was positively correlated with HBsAg level ( r=0.231, P=0.022), and the mRNA expression level was negatively correlated with HBsAg level ( r=-0.245, P=0.014) in patients with HBV infection. Conclusion:There are differences in DNA methylation and mRNA expression of APTR gene in different stages of HBV infection, suggesting that APTR gene may be involved in the immune regulation of HBV infection.
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OBJECTIVE: To investigate the effect of hyperthermia on the expression of PANDAR, LncRNA-p21 and ST8SIA genes in the human lung adenocarcinoma A549 cells. METHODS: A549 cells were randomly divided into 4 groups. The A549 cells in control group were cultured at 37 ℃; the cells at experimental groups were cultured at 40, 42 or 44 ℃ respectively. The cells in these 4 groups were incubated for 1 hour, and the levels of PANDAR, LncRNA-p21 and ST8SIA genes were analyzed by real-time fluorescence quantitative polymerase chain reaction. RESULTS: In cells cultured at 40, 42 or 44 ℃ experimental groups, the relative expression of PANDAR gene was lower than that of control group(P<0.05). In cells cultured at 44 ℃ experimental group, the relative expression of PANDAR gene was lower than that of the 40 and 42 ℃ experimental groups(P<0.05). There was no significant change in the relative expression of LncRNA-p21 and ST8SIA genes among the four groups(P>0.05). CONCLUSION: Hyperthermia decrease the expression of PANDAR gene in A549 cells.
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Objective To investigate the expression of miR?126, miR?355 and exportin?5 in lung cancer. Methods The cancer tissue and the tissue adjacent to carcinoma of 47 cases of patients with lung cancer was used to detect the expression of miR?126, miR?355 and Exportin?5 by the real?time fluorescence quantitative PCR. Results Significant difference of the expression of miR?126 (t=2.02,P=0.03) and exportin?5 (t=4.62,P<0.01) was observed in lung cancer tissue and tissue adjacent to carcinoma. Mature miR?126 and pri?miR?126 (R=0.309 , P = 0.044) had a negative correlation in the tissue adjacent to carcinoma. In the cancer tissue,miR?126 and MRP (R=0.432, P=0.019), miR?335 and k167 (R=0.410, P=0.033) were positively correlated, however, exportin?5 and TOPO (R=0.357, P=0.045), the pri?miR?126 and drinking (R=0.340, P=0.024), the pri?miR?126 and MRP (R=0.427, P=0.027) had a negative correlation relationship. Conclusion Expression of miR?126 and exportin?5 was decreased in lung cancer tissue, which may contribute to the occurrence and development of lung cancer.
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OBJECTIVE: To explore the effect of arsenic on the homeobox D10( HOXD10) gene expression in peripheral blood lymphocytes and human lung adenocarcinoma cell A549. METHODS: ⅰ) A total of 59 workers exposed to arsenic from a arsenic factory were selected as the exposure group and 17 local people without arsenic exposure were chosen as controls by using judgment sampling method. Hydride generation-cold hydrazine trapping-atomic absorption spectrometry was used to detect arsenic levels in urine of these 2 groups. The expression of HOXD10 in peripheral blood lymphocytes was detected by real-time fluorescent quantitative polymerase chain reaction( qRT-PCR).ⅱ) The A549 cells were treated with arsenic trioxide( As_2O_3) with concentration of 0. 0,0. 1,0. 5,1. 0 and 2. 0 μmol/L,and the survival rate of cells was examined by colorimetric assay. The expression of HOXD10 was detected by qRT-PCR. RESULTS: ⅰ) The levels of inorganic arsenic,methylarsonic acid,dimethyl arsenate,total arsenic in the urine,and the relative expression of HOXD10 mRNA in peripheral blood lymphocyte in exposure group were higher than that of the control group( P < 0. 05).ⅱ) As_2O_3 decreased the survival rate of A549 cells in a dose-dependent manner( P < 0. 01) and lead to a dose-dependent increase of HOXD10 mRNA expression( P < 0. 01). A549 cell survival rate and relative expression of HOXD10 mRNA showed a negative correlation,the correlation coefficient was-0. 777( P < 0. 01). CONCLUSION: Arsenic can up-regulate HOXD10 expression in the peripheral blood of occupational arsenic exposure individuals. As_2O_3 can inhibit the proliferation of A549 cells,which may be related to the up-regulation of HOXD10 expression.
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Objective To study the expression characteristics of Let-7 genes in breast cancer.Methods Twenty-eight patients with breast cancer were randomly selected,and their cancer tissue and adjacent normal tissue were collected.TRIzol was used to extract the total RNA and real-time quantitative PCR was used to detect the relative gene expression levels.Results The expression of precursor Let-7 in cancer tissue was (9.65 ± 2.31),which was lower than that in normal tissue (10.05 ± 2.81),P =0.048.Precursor Let-7 had dependence relationship with the long menstruation (b =0.816,P =0.029).The menarche age showed a positive correlation with precursor Let-7 in normal tissue (r =0.502,P =0.048) and a negative correlation in cancer tissue (r =-0.484,P =0.049) Conclusions The expression of precursor Let-7 in cancer tissue is lower than that in normal tissue.The period of menstruation is a protective factor to breast cancer.
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Objective To explore the correlation between serum Lipoxin A4 and clinical grading of chronic hepatitis B patients. Method The serum Lipoxin A4 was detected by Enzyme-Linked Immunosorbent Assay in 94 chronic hepatitis B patients. Results It was found that the level of serum Lipoxin A4 of severe hepatitis patients were significantly lower than mild hepatitis patients and moderate hepatitis patients ( =0.04 and =0.03) . The serum Lipoxin A4 levels were correlated negatively with the ALT and AST levels,respectively =-0.41, =0.019 and R=-0.37,P=0.034. Conclusion These findings support the fact that the serum Lipoxin A4 may contribute to clinical grading of chronic hepatitis B patients.