Fangchinoline Exerts Anticancer Effects on Colorectal Cancer Cells by Evoking Cell Apoptosis via Endoplasmic Reticulum Stress.

We studied the anti-tumor effect of fangchinoline (FAN) against human colorectal cancer cell lines CCL-244 and SW480 and analyzed the mechanism of FAN action. The cell viability and apoptosis were assessed by MTT test and Annexin V-PI staining; caspase-3 activity was measured by Western blotting. The expression of endoplasmic reticulum stress-related proteins was assessed by real-time PCR, Western blotting, and gene transfection. It was found that FAN inhibited cell growth and induced apoptosis in human colorectal cancer cell lines CCL-244 and SW480 in a dose-dependent manner. The caspase-3 inhibitor Ac-DEVD-CHO could reverse the inhibitory effect of FAN. Moreover, FAN significantly increased the expression of endoplasmic reticulum stress-related proteins p-PERK, p-eIF2α, ATF4, and CHOP in CCL-244 and SW480 cells. In addition, endoplasmic reticulum stress inhibitor 4-phenylbutyric acid or CHOP knockdown could prevent FAN-induced apoptosis. Thus, FAN induced apoptosis of human colorectal cancer through activation of endoplasmic reticulum stress.

Meta-analysis of 5-hydroxytryptamine type 2A receptor polymorphisms and migraine susceptibility.

Epidemiologic studies have investigated the association of polymorphisms in 5-hydroxytryptamine type 2A receptor (5HT2A) gene and migraine susceptibility, but the results of those studies are inconclusive. To obtain a more systematic estimation of the association, we conducted a comprehensive search to examine all the eligible studies of 5HT2A polymorphisms and migraine risk. The odd ratios (ORs) with 95% confidence intervals (CIs) were used to determine the strength of the association. Publication bias was analyzed by Begg's funnel plots. Seven eligible studies regarding 5HT2A T102C and A-1438G polymorphisms with 721 cases and 713 controls were included in this meta-analysis. Overall, no significant associations were found between 5HT2A T102C (for T vs. C: OR = 1.029, 95% CI = 0.870-1.217, p = 0.739; for TT vs. CC: OR = 1.083, 95% CI = 0.760-1.544, p = 0.657; for TT + TC vs. CC: OR = 1.066, 95% CI = 0.802-1.416, p = 0.662; for TT vs. TC + CC: OR = 1.017, 95% CI = 0.774-1.336, p = 0.904) or A-1438G (for T vs. C: OR = 0.996, 95% CI = 0.726-1.365, p = 0.979; for TT vs. CC: OR = 0.983, 95% CI = 0.511-1.891, p = 0.960; for TT + TC vs. CC: OR = 1.118, 95% CI = 0.654-1.910, p = 0.684; for TT vs. TC + CC: OR = 0.890, 95% CI = 0.528-1.499, p = 0.661) polymorphisms and migraine risk. The further subgroup analysis by ethnicity, assay and disease type also found no significant association using four genetic models. Meanwhile, the publication bias analysis suggests that there is no publication bias in these studies. In conclusion, our current meta-analysis implies that 5HT2A T102C and A-1438G polymorphisms may be not risk factors in the pathogenesis of migraine.

miR-371b-5p promotes cell proliferation, migration and invasion in non-small cell lung cancer via SCAI.

OBJECTIVE: Multiple gene targets have been reported for treatment of non-small cell lung cancer (NSCLC), however, the accompanying genetic tolerance was reported increasingly. Therefore, it is important to find new biomarkers or therapeutic targets in treatment of NSCLC. METHODS: The expression levels of miR-371b-5p were detected by qRT-PCR in NSCLC tissues and cell lines. To evaluate the effect of miR-371b-5p on NSCLC progression, we first transfected the miR-371b-5p inhibitor for construction of the miR-371b-5p down-regulated cell model. Then the cell proliferation, migration, invasion and cell apoptosis were detected. In addition, the expression levels of adhesion factors were detected. The target gene of miR-371b-5p was identified by bioinformatics analysis, and rescue experiment was conducted to validate the effect of miR-371b-5p on proliferation, migration and invasion of NSCLC. RESULTS: Our findings revealed that the miR-371b-5p was overexpressed in NSCLC and could markedly promote the cell proliferation, migration and invasion. Expression levels of both intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were significantly down-regulated when treated by miR-371b-5p inhibitor. Moreover, dual-luciferase reporter assay showed that the miR-371b-5p targeted SCAI in regulation of cell proliferation, migration and invasion, and the expression of miR-371b-5p was negatively associated with SCAI in NSCLC tissues and cell lines. Rescue experiment revealed that the miR-371b-5p could rescue the effect of SCAI on cell proliferation, migration and invasion. CONCLUSION: Our results suggest that the miR-371b-5p and SCAI may serve as novel prognostic biomarkers and therapeutic targets for NSCLC.

Fangchinoline inhibits the proliferation of SPC-A-1 lung cancer cells by blocking cell cycle progression.

Fangchinoline (Fan) is a bioactive compound isolated from the Chinese herb Stephania tetrandra S. Moore (Fen Fang Ji). The aim of the present study was to investigate the effect of Fan on the proliferation of SPC-A-1 lung cancer cells, and to define the associated molecular mechanisms. Following treatment with Fan, Cell Counting Kit-8, phase contrast imaging and Giemsa staining assays were used to detect cell viability; flow cytometry was performed to analyze the cell cycle distribution; and reverse transcription-quantitative polymerase chain reaction and western blot assays were used to investigate changes in the expression levels of cell cycle-associated genes and proteins. In the present study, treatment with Fan markedly inhibited the proliferation of SPC-A-1 lung cancer cells and significantly increased the percentage of cells in the G0/G1 phase of the cell cycle in a dose-dependent manner (P<0.05 for 2.5-5 µm; P<0.01 for 10 µm), whereas the percentage of cells in the S and G2/M phases were significantly reduced following treatment (P<0.05 for 5 µm; P<0.01 for 10 µm). Mechanistically, Fan significantly reduced the mRNA expression levels of cyclin D1, cyclin-dependent kinase 4 (CDK4) and CDK6 (P<0.05 for 2.5-5 µm; P<0.01 for 10 µm), which are key genes in the regulation of the G0/G1 phase of the cell cycle. Furthermore, treatment with Fan also decreased the expression of phosphorylated retinoblastoma (Rb) and E2F transcription factor-1 (E2F-1) proteins (P<0.05 for 5 µm; P<0.01 for 10 µm). In summary, the present study demonstrated that Fan inhibited the proliferation of SPC-A-1 lung cancer cells and induced cell cycle arrest at the G0/G1 phase. These effects may be mediated by the downregulation of cellular CDK4, CDK6 and cyclin D1 levels, thus leading to hypophosphorylation of Rb and subsequent suppression of E2F-1 activity. Therefore, the present results suggest that Fan may be a potential drug candidate for the prevention of lung cancer.

Tacrolimus (FK506) prevents early retinal neovascularization in streptozotocin-induced diabetic mice.

Diabetic retinopathy is a complex disease that has potential involvement of inflammatory in its pathogenesis. We hypothesized that tacrolimus (FK506), one of the potent immunosuppressive agent, could be effective against diabetic retinopathy, which involves significant inflammation. The aim of the present study was to investigate the effects of FK506 in early retinal changes of streptozotocin-induced diabetic mice. The effect of FK506 treatment (10 µg per eye for one week) was evaluated by TNF-a, VEGF, iNOS and COX-2 protein levels measurement, neovascularization, and the activation of NF-kB in the retina. Increased amounts of cytokines, neovascularization, inflammatory markers and activation of NF-kB were observed in retina from diabetic mice. FK506 treatment significantly lowered retinal TNF-a, VEGF, iNOS and COX-2. Further, treatment with FK506 significantly suppressed diabetes-related neovascularization, as well as the activation of NF-kB. These data demonstrated that FK506 attenuates the degree of retinal inflammation and preserving the neovascularization in early diabetic mice.