MicroRNA 375 regulates proliferation and migration of colon cancer cells by suppressing the CTGF-EGFR signaling pathway.

MicroRNA 375 (MIR375) is significantly down regulated in human colorectal cancer (CRC) tissues; we have previously identified MIR375 as a colon cancer associated microRNA (miRNA). We identified putative MIR375 target genes by comparing the mRNA microarray analysis data of MIR375-overexpressing cells with the candidate MIR375 target genes predicted by public bioinformatic tools. We investigated that the connective tissue growth factor (CTGF) is a direct target gene of MIR375. Expression of CTGF, a ligand of epidermal growth factor receptor (EGFR), was markedly enhanced in human CRC tissues in comparison with the corresponding normal colon tissues. We demonstrated that the expression levels of molecules in EGFR signaling pathways were regulated by MIR375 in colorectal cells. Using immunohistochemistry and the xenograft of MIR375-overexpressing colorectal cells in mice, we showed that MIR375 regulates cell growth and proliferation, angiogenesis, cell migration, cell cycle arrest, apoptosis, and necrosis in colon cells. Furthermore, results of MIR375 overexpression and cetuximab treatment indicated that the apoptosis and necrosis in colon cells were synergistically enhanced. Our results suggest that the down-regulation of MIR375 modulates EGFR signaling pathways in human colorectal cells and tissues by increasing CTGF expression; therefore, MIR375 may have a therapeutic value in relation to human CRC.

Knowledge, attitude and prevention practices of garment factory workers regarding the largest Dengue outbreak on record in Bangladesh.

The aim of this cross-sectional study was to assess the knowledge, attitudes and prevention practices (KAP) among the garment factory worker population in Bangladesh regarding a historical dengue outbreak in 2019. A total of 400 participants were selected by simple random sampling, and questionnaire-based interviews were conducted. The average score of knowledge, attitude and prevention practice was 8.33 ± 2.35, 6.32 ± 1.20 and 6.31 ± 1.50, respectively. Only 76 out of 400 participants (19%) scored above 10 (all university-educated). Participant workers reported both negative and positive attitudes regarding dengue fever (DF). Negative attitudes included an expectation of increased mortality and strained family finances from DF attacks. A significantly high number of participants (92%) believed that death from DF was inevitable. Positive attitudes included optimism about DF eradication potentials and eagerness to help and donate blood to sick relatives. Participants primarily learned about the DF prevention from mass media (244/400; 61.0%) and social media (97/400; 24.25%). The most popular prevention measures adopted were mosquito repellent incense (344/400; 86.0%) and mosquito nets (389/400; 97.25%). While most participants (358/400; 89.5%) cleaned areas where mosquitos lay eggs, only 169 out of 400 (42.25%) regularly treated with chemical sprays. Only 182 out of 400 (45.5%) reported receiving DF prevention training in the factory. Correlation between DF knowledge and education was statistically significant (r = .38, p < .01, n = 398). Correlation between DF knowledge and work experience was insignificant (r = .01, p > .01, n = 398). Age and DF knowledge were not correlated (r = 0.07, p > .01, n = 398). In conclusion, gaps in KAP for dengue could be addressed by government-sponsored educational programmes that utilize the power of mass/social media for dengue prevention and control. More KAP surveillance studies are needed for other sectors of the society.

MicroRNA 429 Regulates Mucin Gene Expression and Secretion in Murine Model of Colitis.

BACKGROUND AND AIMS: miRNAs are non-coding RNAs that play important roles in the pathogenesis of human diseases by regulating target gene expression in specific cells or tissues. We aimed to detect miRNAs related to ulcerative colitis [UC], identify their target molecules, and analyse the correlation between the miRNAs and their target genes in colorectal cells and dextran sulphate sodium [DSS]-induced mouse colitis. METHODS: UC-associated miRNAs were identified by miRNA microarray analysis using DSS-induced colitis and normal colon tissues. The results were validated by quantitative real-time polymerase chain reaction [RT-PCR]. We identified target genes of MIR429, a colitis-associated miRNA, from our screen by comparing the mRNA microarray analysis in MIR429-overexpressed cells with predicted candidate target genes. We constructed luciferase reporter plasmids to confirm the effect of MIR429 on target gene expression. The protein expression of the target genes was measured by western blot,enzyme-linked immunosorbent assay [ELISA] analysis, or immunohistochemistry. RESULTS: We identified 37 DSS-induced colitis associated miRNAs. We investigated MIR429 that is down-regulated in DSS-induced colitis, and identified 41 target genes of MIR429. We show that the myristoylated alanine-rich protein kinase C substrate [MARCKS] is a direct target of MIR429. MARCKS mRNA and protein expression levels are down-regulated by MIR429, and MIR429 regulates the expression of MARCKS and MARCKS-mediated mucin secretion in colorectal cells and DSS-induced colitis. In addition, anti-MIR429 up-regulates MARCKS expression in colorectal cell lines. CONCLUSION: Our findings suggest that MIR429 modulates mucin secretion in human colorectal cells and mouse colitis tissues by up-regulating of MARCKS expression, thereby making MIR429 a candidate for anti-colitis therapy in human UC.

MicroRNA 196B regulates FAS-mediated apoptosis in colorectal cancer cells.

Using miRNA microarray analysis, we identified 31 miRNAs that were significantly up-regulated or down-regulated in colon cancer tissues. We chose MIR196B, which was specifically up-regulated in colon cancer, for further study. We identified 18 putative MIR196B target genes by comparing between the mRNAs down-regulated in MIR196B-overexpressed cells and the assumed MIR196B target genes predicted by public bioinformatics tools. The association between MIR196B and FAS was verified in this study. FAS expression was constitutively elevated in normal human colorectal tissues. However, its expression was often reduced in human colorectal cancer. The decrease in FAS expression could be responsible for the reduction of apoptosis in colorectal cancer cells. In colorectal cancer tissue, we showed that MIR196B up-regulation was mutually followed by down regulation of FAS expression. We also showed that MIR196B directly repressed FAS expression in colorectal cells. Furthermore, anti-MIR196B up-regulated FAS expression and increased apoptosis in colorectal cancer cell lines. Our results suggest that the up-regulation of MIR196B modulates apoptosis in colorectal cancer cells by partially repressing FAS expression and that anti-MIR196B could be a potential candidate as an anti-cancer drug in colorectal cancer therapy.