Effects of lncRNA MEG3 on proliferation and apoptosis of gallbladder cancer cells through regulating NF-κB signaling pathway.

OBJECTIVE: To explore the effects of long non-coding ribonucleic acid (lncRNA) maternally expressed gene 3 (MEG3) on proliferation and apoptosis of gallbladder cancer (GBC) cells and its molecular mechanism. MATERIALS AND METHODS: The relative expression level of lncRNA MEG3 in GBC cell lines was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The lncRNA MEG3 expression plasmids were constructed, the cell proliferation ability was detected via Cell Counting Kit-8 (CCK-8) assay and colony formation assay, and the apoptosis was detected via flow cytometry. The effects of lncRNA MEG3 expression on endoplasmic reticulum stress (ERS)-related proteins were determined using Western blotting, and the changes in nuclear factor-κB (NF-κB) protein in the nucleus were determined after overexpression of lncRNA MEG3. RESULTS: The expression of lncRNA MEG3 in three kinds of GBC cell lines was lower than that in human immortalized normal biliary epithelial cells (p<0.05). The results of CCK-8 assay and colony formation assay showed that overexpression of lncRNA MEG3 significantly reduced the proliferation rate and colony formation ability of GBC-SD cells compared with negative control (NC) group (p<0.05, p<0.05). According to the results of flow cytometry, the apoptosis rate was higher in lncRNA MEG63 overexpression group compared with that in NC group (p<0.05). Moreover, the ERS-related proteins (MANF, GRP78, and caspase-3) were remarkably upregulated in lncRNA MEG63 overexpression group compared with those in NC group, indicating that ERS is activated by lncRNA MEG63 overexpression. The NF-κB signal in GBC cells was activated by lncRNA MEG3. CONCLUSIONS: LncRNA MEG3 activates the NF-κB signal in GBC cells to affect the proliferation and apoptosis of GBC cells.

Linc00707 promotes cell proliferation, invasion, and migration via the miR-30c/CTHRC1 regulatory loop in breast cancer.

OBJECTIVE: Breast cancer (BC) is the most common malignant tumor in women. We aimed at investigating the function of long non-coding RNA LINC00707 in BC and the potential mechanism. PATIENTS AND METHODS: The expression level of linc00707 was determined using the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) in BC tissues and cell lines. The Cell Counting Kit-8 (CCK-8) and colony formation assays were performed to detect the potential influence of LINC0070 on the proliferation ability of the BC cells. Also, the invasion and migration abilities were assessed by the transwell assay. Furthermore, with the bioinformatic analysis and the Dual-Luciferase Reporter Gene Assay, we analyzed the interaction in LINC00707/miR-30c/CTHRC1 regulatory loop. The regulatory effects of LINC00707/miR-30c/CTHRC1 on BC were finally determined. RESULTS: LINC00707 was significantly upregulated in BC tissues and cell lines. The knockdown of LINC00707 inhibited proliferation, invasion, and migration in MDA-MB-231 cells, while the overexpression of LINC00707 achieved the opposite results in MDA-MB-468 cells. LINC00707, acting as a competing endogenous RNA (ceRNA), could sponge miR-30c to upregulate CTHRC1, thus promoting BC progression. CONCLUSIONS: LINC00707 was highly expressed in BC tissues and cells. It promoted cell proliferation, invasion, and migration via miR-30c/CTHRC1 regulatory loop. This might provide a novel target for the diagnosis, treatment, and prognosis for BC.

TGF-ß1 promotes pulmonary arterial hypertension in rats via activating RhoA/ROCK signaling pathway.

OBJECTIVE: To investigate the influence of the Rho/Rho-associated kinase (ROCK) signaling pathway in rats with pulmonary arterial hypertension (PAH) under the intervention with transforming growth factor-beta 1 (TGF-ß1). MATERIALS AND METHODS: A total of 30 rats were divided into three groups using a random number table, including control group (healthy rats, n=10), model group (PAH rats, n=10), and TGF group (PAH rats injected with 5 ng/mL TGF-ß1 recombinant protein, n=10). The systolic blood pressure, ventricular hypertrophy index, pathological changes in lung tissues, TGF-ß1 level, protein, and messenger ribonucleic acid (mRNA) expressions of RhoA and ROCK, as well as concentrations of serum nitric oxide (NO) and endothelin-1 (ET-1) were detected via hemodynamics test, hematoxylin and eosin (HE) staining, immunohistochemical method, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA). RESULTS: The results of hemodynamics test showed that the right ventricular systolic pressure was increased markedly in model group (46.53±8.81) and TGF group (56.79±9.12) compared with that in control group (26.03±4.21) (p<0.05). The mean pulmonary systolic pressure in model group (25.89±1.92) and TGF group (29.41±1.91) was evidently higher than that in control group (15.77±2.71) (p<0.05). According to the results of heart weight measurement, model group (0.5118±0.1635) exhibited a higher ventricular hypertrophy index than control group (0.2908±0.0313) (p<0.05) but a lower ventricular hypertrophy index than TGF group (0.7231±0.1004) (p<0.05). The medial thickness of the pulmonary artery of the rats was observed through the HE staining. It was found that compared with control group, the medial thickness of the pulmonary artery was increased significantly in model group (p<0.05), while it was raised more prominently in TGF group, higher than that in model group, suggesting that TGF-ß1 expression can increase the medial thickness of the pulmonary artery. It was manifested in immunohistochemical results that the protein expression of RhoA in the left lung tissues rose notably in model group compared with that in control group (p<0.05), and it was also raised remarkably in TGF group in comparison with that in model group (p<0.05), illustrating that the protein expression of TGF can activate the activity of RhoA and ROCK. The results of RT-PCR indicated that the mRNA expressions of RhoA and ROCK in the left lung tissues were elevated distinctly in model group and TGF group compared with those in control group (p<0.05), and the increases were more apparent in TGF group than those in model group (p<0.05). It was revealed in ELISA results that in comparison with control group, model group, and TGF group had markedly increased concentrations of serum NO and ET-1 (p<0.05), while the rises of serum NO and ET-1 concentrations in TGF group were the most prominent compared with those in model group (p<0.05). CONCLUSIONS: Overexpressed TGF-ß1 can activate the RhoA/ROCK signaling pathway, thus promoting the occurrence and development of PAH.

LINC00511 can promote the proliferation, migration and invasion of esophageal cancer cells through regulating microRNA-150-5p.

OBJECTIVE: The aim of this study was to investigate the potential role of LINC00511 in esophageal cancer (ECa), and to explore its underlying mechanism through in vitro cell experiments. PATIENTS AND METHODS: LINC00511 expression in ECa was analyzed by GEPIA database and verified by real-time fluorescence quantitative polymerase chain reaction (qPCR). The bioinformatics website was used to analyze the miRNAs that can bind to LINC00511, and the regulatory relationship between them was verified through Luciferase assay, qPCR as well as Western blotting analysis. Then, the impacts of LINC00511 and microRNA-150-5p on the proliferation or invasiveness of ECa cell lines Kyse30 and ECA109 were investigated by cell counting kit-8 (CCK-8) test and transwell experiment, respectively. Meanwhile, cell cycle and apoptosis were detected by flow cytometry. RESULTS: Analysis results of the GEPIA database revealed that LINC00511 had a significant high expression in ECa tissue samples in comparison with normal control ones, which is consistent with qPCR results. Meanwhile, a significant negative correlation was found between LINC00511 and microRNA-150-5p. In brief, LINC00511 was able to bind to microRNA-150-5p and inhibited its expression. Besides, overexpression of LINC00511 enhanced ECa cell proliferation and migration, accelerated cell cycle, and suppressed cell apoptosis, while transfection with microRNA-150-5p mimics caused the opposite effects. CONCLUSIONS: This study shows for the first time that LINC00511 modulates the progression of ECa by binding to microRNA-150-5p.

[Clinical observation on the treatment of ischemic heart disease with Astragalus membranaceus].

92 patients suffering from ischemic heart disease was successfully treated with Astragalus membranaceus (AM). The effect of the treatment was compared with that of Nifedipine and Tab. Salviae miltiorrhizae. The clinical practice showed that the group treated with AM yielded better results. After having administered the drug, the patients were markedly relieved from angina pectoris (heart stroke). Meanwhile the improvement of clinical objective index such as electrocardiogram (EKG) and impedance cardiogram can also be observed. The effective rate of EKG improvement was 82.6%. The treatment of ischemic heart disease with AM was significantly more effective in comparing with control group (P < 0.05).