microRNA-142-3p inhibits apoptosis and inflammation induced by bleomycin through down-regulation of Cox-2 in MLE-12 cells.

microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 µg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 µg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.

microRNA-142-3p inhibits apoptosis and inflammation induced by bleomycin through down-regulation of Cox-2 in MLE-12 cells

microRNA (miR)-142-3p is implicated in malignancy and has been identified as a biomarker for aggressive and recurrent lung adenocarcinomas. This study aimed to evaluate the inhibitory effect of miR-142-3p on apoptosis and inflammation induced by bleomycin in MLE-12 cells. MLE-12 cells were first transfected either with miR-142-3p mimic or miR-142-3p inhibitor and then the cells were exposed to 50 μg/mL of bleomycin. Thereafter, cell viability, apoptosis and the expression of pro-inflammatory cytokines were assessed using CCK-8, flow cytometry, RT-PCR and western blot analyses. Cox-2, PI3K, AKT and mTOR expressions were detected by western blotting after bleomycin was administered together with NS-398 (an inhibitor of Cox-2). As a result, cell viability was significantly decreased, as well as apoptosis and the expression of IL-1 and TNF-α were remarkably increased after 50 and 100 μg/mL of bleomycin administration. miR-142-3p overexpression alleviated bleomycin-induced apoptosis and overproduction of these two pro-inflammatory cytokines, while miR-142-3p suppression exhibited completely opposite results. Up-regulation of Cox-2 and inactivation of PI3K/AKT/mTOR were found in bleomycin-pretreated cells, while these abnormal regulations were partially abolished by miR-142-3p overexpression and NS-398. In conclusion, this study demonstrated that miR-142-3p overexpression protected bleomycin-induced injury in lung epithelial MLE-12 cells, possibly via regulating Cox-2 expression and PI3K/AKT/mTOR signaling pathway. These findings provide evidence that miR-142-3p may be a therapeutic strategy for idiopathic pulmonary fibrosis (IPF) treatment.

Effect of CDH1 gene methylation on transforming growth factor (TGF-ß)-induced epithelial-mesenchymal transition in alveolar epithelial cell line A549.

The aim of this study was to investigate the specific molecular mechanism of the transforming growth factor ß (TGF-ß)-induced epithelial-mesenchymal transition in a lung cancer cell line, and to provide new ideas for targeting therapy of lung cancer. A549 cells were treated with different concentrations of TGF-ß and 5-aza-deoxycytidine (5-aza-dC). The morphological changes after the intervention were observed. The change in the expression of the epithelial marker E-cadherin (E-cad) was detected by Western blot. The proliferation of A549 cells was measured using the MTT assay. Cell movement and invasion capacity was evaluated with the cell scratch test and invasion test. TGF-ß induced A549 cells to transform to a mesenchymal cell morphology and downregulated the expression of E-cad, and 5-aza-dC inhibited this phenomenon. Compared with the control group, the number of transmembrane cells was higher and cell migration was markedly increased in the experimental group with continued culture in the presence of 10 ng/mL TGF-ß, showing significant differences (P < 0.05). CDH1 gene methylation is involved in TGF-ß-induced epithelial-mesenchymal transition in the alveolar epithelial cell line A549.

Contraceptive effect of intrauterine application of Lugol's solution.

PIP: The contraceptive effect of local application of Lugol's solution (5% elemental iodine and 10% potassium iodide in aqueous solution) to the uterine cavity was evaluated in rats, 1 monkey and humans. 18 experimental rats received .1 ml Lugol's solution injected into the left uterine horn to cover the entire cavity; 2 received .05 ml solution covering two-thirds of the left uterine horn; 12 control rats received saline in the left horn; right uterine horns were left intact. Control rats had 76 pregnancies in the left horn and 70 in the right; the 18 rats treated with .1 ml solution had no pregnancies in the left horn and 121 in the right; 2 rats receiving .05 ml solution had 2 and 3 pregnancies, respectively, in the left horn. Lugol's solution prevented nidation in rats, and its action was local rather than systemic. Lugol's solution applied transvaginally to the uterine cavity of 1 Macaca mulatta adult virgin female monkey after confirmed existence of a developed Graffian follicle was followed by artificial insemination in the treated cycle and 2 cycles later. Ovulation and menstruation occurred during the treated and subsequent cycles, but conception occurred in the third cycle. Effect of Lugol's solution appears temporary. Preliminary clinical observation in a limited number of human volunteers in the U.S., Japan and Mexico who had Lugol's solution applied to the uterine cavity using a cotton-tip applicator indicates that application on Days 16 and 17 of the cycle resulted in no contraceptive effect, but on Day 20-24, resulted in no pregnancies through contraceptive or abortive effect. No irregular and/or abnormal bleeding occurred. A long-term observation and evaluation of the study is warranted to determine side effects of monthly solution application, such as endometrial changes and irregular bleeding. This procedure may be useful in cases of rape because of the endometrial effect and direct spermicidal effect of Lugol's solution if applied soon after coitus. A discussion of study limitations is presented, and the need for development of a simple, harmless and inexpensive contraceptive method which can be practiced by paramedical personnel as well as physicians is stressed.^ieng