11 research outputs found
MiR-145 expression accelerates esophageal adenocarcinoma progression by enhancing cell invasion and anoikis resistance.
No full textBACKGROUND: 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. METHODS: 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. RESULTS: 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. DISCUSSION: 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
The role of miRNA-145 in esophageal adenocarcinoma.
No full text 46 Background: Carcinoma of the esophagus has become one of the fastest growing solid tumors in the world over the past 20 years and the 6th most common cause of death in the world. We previously conducted a study to profile the expression of miRNAs in esophageal adenocarcinoma (EAC) pre and post induction therapy. Out of the miRNAs discovered, miR-145, a known tumor suppressor miRNA, was upregulated 8-fold upon induction therapy however its expression was associated with shorter disease-free survival. This unexpected result was explored. Methods: In order to study the role of miR-145 in EAC, miRNA-145 was overexpressed in the EAC cell line OE33 (OE33 miR145). After validation of the expression of miR-145, several hallmarks of cancer such as cell proliferation, resistance to apoptosis and cell adhesion were analyzed. Results: There was no difference in cell proliferation and resistance to radiation between OE33 miR145 and OE33 control. However, there was a significant difference in cell adhesion. OE33 miR145 cells reattach faster than the OE33 control (72.3% cells reattached against 40.5% cells reattached after 3h, n=3, p<0.05). Furthermore, OE33 miR145 cells are able to recover better than OE33 control cells after 72 hours culture in suspension (126.5 colonies against 63.5 colonies after 5 days of culture, n=2, p<0.05). Conclusions: While there was no difference between the two OE33 populations in an attached state, clear differences appeared when cells were in a detached state. Although more work is required to confirm this effect, expression of miR-145 could help EAC cells in circulation by protecting them against apoptosis and helping them to reattach faster which could in turn facilitate distant metastasis. </jats:p
MiR-145 inhibited cell proliferation, delayed wound closure and enhanced anoikis in ESCC cells.
No full text<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 expression did not affect EAC cells proliferation.
No full text<p>Cell proliferation assay of the pcmv and miR-145 EAC cell lines, n = 3</p
MiR-145 enhanced cell adhesion to fibronectin but not vitronectin.
No full text<p>OE33 (A), SK-GT-4 (B) and FLO-1 (C) were plated onto plates pre-treated with either fibronectin or vitronectin. After 30 mins, the detached cells were counted. *: p<0.05, n = 3.</p
MiR-145 accelerated wound closure and enhanced cell invasion.
No full text<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 did not generally affect the EAC resistance to chemotherapy drugs.
No full text<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 protected EAC cells against anoikis.
No full text<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 enhanced the clonogenic potential of OE33 and SK-GT-4 after suspension culture.
No full text<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
