MiR-145 Expression Accelerates Esophageal Adenocarcinoma Progression by Enhancing Cell Invasion and Anoikis Resistance

<div><p>Background</p><p>Carcinoma of the esophagus has a high case fatality ratio and is now the 6th most common cause of cancer deaths in the world. We previously conducted a study to profile the expression of miRNAs in esophageal adenocarcinoma (EAC) pre and post induction therapy. Of the miRNAs differentially expressed post induction chemoradiation, miR-145, a known tumor suppressor miRNA, was upregulated 8-fold following induction therapy, however, its expression was associated with shorter disease-free survival. This unexpected result was explored in this current study.</p><p>Methods</p><p>In order to study the role of miR-145 in EAC, miRNA-145 was overexpressed in 3 EAC cell lines (OE33, FLO-1, SK-GT-4) and one ESCC cell line (KYSE-410). After validation of the expression of miR-145, hallmarks of cancer such as cell proliferation, resistance to chemotherapy drugs or anoikis, and cell invasion were analyzed.</p><p>Results</p><p>There were no differences in cell proliferation and 5 FU resistance between miR145 cell lines and the control cell lines. miR-145 expression also had no effect on cisplatin resistance in two of three cell lines (OE33 and FLO-1), but miR-145 appeared to protect SK-GT-4 cells against cisplatin treatment. However, there was a significant difference in cell invasion, cell adhesion and resistance to anoikis. All three EAC miR-145 cell lines invaded more than their respective controls. Similarly, OE33 and SK-GT-4 miR-145 cell lines were able to survive longer in a suspension state.</p><p>Discussion</p><p>While expression of miR-145 in ESCC stopped proliferation and invasion, expression of miR-145 in EAC cells enhanced invasion and anoikis resistance. Although more work is required to understand how miR-145 conveys these effects, expression of miR-145 appears to promote EAC progression by enhancing invasion and protection against anoikis, which could in turn facilitate distant metastasis.</p></div

MiR-145 inhibited cell proliferation, delayed wound closure and enhanced anoikis in ESCC cells.

<p>(A) Cell proliferation and (B) wound healing assay of KYSE-410 pcmv and KYSE-410 miR-145 cells. miR-145 expression in KYSE-410 led to decreased numbers of colonies after cell suspension culture (C) and enhanced PARP and caspase 3 cleavage (D).*: p<0.05, n = 3.</p

MiR-145 protected EAC cells against anoikis.

<p>pcmv and miR-145 EAC cell lines were cultured in suspension for 72 h. The levels of cleaved PARP and cleaved caspase 3 were assessed by Western Blotting.</p

MiR-145 expression did not affect EAC cells proliferation.

<p>Cell proliferation assay of the pcmv and miR-145 EAC cell lines, n = 3</p

MiR-145 did not generally affect the EAC resistance to chemotherapy drugs.

<p>OE33, FLO-1 and SK-GT-4 (pcmv and miR-145) cells were cultured for 72 h with either (A) cisplatin (5 µM) or (B) 5-fluorouracil (35 µM) and their respective control. Live cell number was then assessed. Results show the percentage of live cells compared to the control treatment. *: p<0.05, n = 3.</p

MiR-145 accelerated wound closure and enhanced cell invasion.

<p>Wound healing assay (A) with pcmv and miR-145 cell lines. The wound length was assessed after 8 h culture. *: p<0.05, n = 3. Cell invasion assay with pcmv and miR-145 cell lines (B). *: p<0.05, n = 3.</p

MiR-145 enhanced the clonogenic potential of OE33 and SK-GT-4 after suspension culture.

<p>Photo images of clonogenic assay after 72 h suspension culture with OE33 and SK-GT-4 (A). Results showed the average percentage of colonies formed after suspension culture compare to the number of colonies formed in monolayer (B). *: p<0.05, n = 2.</p

Association between UGT1A1*28 Polymorphisms and Clinical Outcomes of Irinotecan-Based Chemotherapies in Colorectal Cancer: A Meta-Analysis in Caucasians

<div><p>Background</p><p>Whether UGT1A1*28 genotype is associated with clinical outcomes of irinotecan (IRI)-based chemotherapy in Colorectal cancer (CRC) is an important gap in existing knowledge to inform clinical utility. Published data on the association between UGT1A1*28 gene polymorphisms and clinical outcomes of IRI-based chemotherapy in CRC were inconsistent.</p> <p>Methodology/Principal Findings</p><p>Literature retrieval, trials selection and assessment, data collection, and statistical analysis were performed according to the PRISMA guidelines. Primary outcomes included therapeutic response (TR), progression-free survival (PFS) and overall survival (OS). We calculated odds ratios (OR) and hazard ratios (HR) with 95% confidence intervals (CI). Twelve clinical trials were included. No statistical heterogeneity was detected in analyses of all studies and for each subgroup. Differences in TR, PFS and OS for any genotype comparison, UGT1A1*28/*28 versus (vs) UGT1A1*1/*1 (homozygous model), UGT1A1*1/*28 vs UGT1A1*1/*1 (heterozygous model), and UGT1A1*28/*28 vs all others (recessive model, only for TR) were not statistically significant. IRI dose also did not impact upon TR and PFS differences between UGT1A1 genotype groups. A statistically significant increase in the hazard of death was found in Low IRI subgroup of the homozygous model (HR = 1.48, 95% CI = 1.06–2.07; P = 0.02). The UGT1A1*28 allele was associated with a trend of increase in the hazard of death in two models (homozygous model: HR = 1.22, 95% CI = 0.99–1.51; heterozygous model: HR = 1.13, 95% CI = 0.96–1.32). These latter findings were driven primarily by one single large study (Shulman et al. 2011).</p> <p>Conclusions/Significance</p><p>UGT1A1*28 polymorphism cannot be considered as a reliable predictor of TR and PFS in CRC patients treated with IRI-based chemotherapy. The OS relationship with UGT1A1*28 in the patients with lower-dose IRI chemotherapy requires further validation.</p> </div

Flow diagram for study selection in meta-analysis.

<p>Flow diagram for study selection in meta-analysis.</p

Raw data and analytical code Turissini et al. 2015

Raw data and analytical code Turissini et al. 201