Microrna-143 (miR-143) has been suggested to be a tumor suppressor, yet its role in hematological tumors has not been determined. Thus, we aimed to explore the expression and function of miR-143 in leukemia cells. miR-143 expression was assessed in bone marrow samples from 63 leukemia patients and 15 healthy controls using q-PCR, and its correlation with DNMT3A expression was determined. In addition, after lentiviral-mediated miR-143 overexpression, K562 cell proliferation was evaluated using CCK-8 analysis; cell cycle progression and apoptosis were determined using flow cytometry. The expression of Bcl-2 and pro-caspase-3 and -9 was assessed by q-PCR and western blot analysis, respectively. Leukemia patients had significantly lower relative miR-143 expression than healthy controls (P=0.004), and the expression levels of miR143 and DNMTA3A were negatively correlated (r=-0.663, P=0.001). Overexpression of miR-143 decreased DNMT3A mRNA and protein expression, and significantly reduced K562 cell proliferation at 72 and 96 h (both P ≤ 0.018). In addition, reduced colony formation and cell cycle progression were observed upon miR-143 overexpression. Flow cytometric analysis revealed that the early apoptosis rate was higher in the miR-143 group than the rate in the NC group. Bcl-2 mRNA expression and pro-caspase-3 and -9 protein expression were reduced in the miR-143-expressing cells. These findings suggest that miR-143 plays an important role in leukemia cell proliferation and apoptosis, possibly through silencing of DNMT3A. Further studies are necessary to determine the prognostic value and therapeutic potential of targeting miR-143.

https://www.spandidos-publications.com/10.3892/or.2014.3078/download

Overexpression of microRNA-143 inhibits growth and induces apoptosis in human leukemia cells

AIM: To study the characteristics of APC (adenomatous polyposis coli) gene germline mutation in Chinese patients with familial adenomatous polyposis (FAP).

METHODS: APC gene from 14 FAP families was amplified by polymerase chain reaction (PCR) and underwent direct sequencing to determine the micromutation type. For the samples without micromutation, the large fragment deletion of APC gene was examined by multiplex ligation-dependent probe amplification (MLPA).

RESULTS: There were gene micromutations in 9 families with a micromutation detection rate of 64.3% (9/14), including 6 frameshift mutations (66.7%), 1 nonsense mutation (11.1%) and 2 splicing mutations (22.2%). Large fragment deletions were detected by MLPA in 2 families. The total mutation detection rate of micromutations and large fragment deletions was 78.6% (11/14).

CONCLUSION: The detection rate of APC gene germline mutation can be improved by direct sequencing combined with MLPA large fragment deletion detection.

APC gene mutations in Chinese familial adenomatous polyposis patients.

Altered expression of miRNAs contributes to development and progression of non-small cell lung cancer (NSCLC), while transforming growth factor-β (TGF-β) promotes NSCLC cell epithelial-mesenchymal transition. This study aimed to investigate the effects of TGF-β-induced miR‑143 expression in regulation of NSCLC cell viability, invasion capacity in vitro, and xenograft formation and growth in nude mice. NSCLC A549 cells treated with TGF-β were subjected to miRNA microarray analysis and miR‑143 was selected for further study of tumor cell viability, wound healing, invasion capacity in vitro, and tumor growth in nude mice. TGF-β treatment upregulated expression of 16 miRNAs and downregulated expression of 42 miRNAs in A549 cells. qRT-PCR and in situ hybridization data showed that miR‑143 was significantly downregulated in 24 NSCLC and lymph node metastatic tumor tissues, but upregulated by TGF-β treatment in A549 cells. In vitro experiments showed that miR‑143 expression could significantly suppress NSCLC cell viability and invasion capacity, and nude mouse experiments confirmed the in vitro data. Bioinformatic data predicted that Smad3, CD44 and K-Ras were the targeting genes of miR‑143. TGF-β-induced miR‑143 expression was associated with suppressed expression of Smad3, CD44, and K-Ras. This study sheds light on the role of TGF-β in upregulation of miR‑143 and the role of miR‑143 in NSCLC progression, indicating that the target of miR‑143 expression could be further studied as a novel therapeutic strategy for future control of NSCLC.

/pdf/transforming-growth-factor-b-induced-mir-143-expression-in-2cpdixkiw8.pdf

Transforming growth factor-β-induced miR‑143 expression in regulation of non-small cell lung cancer cell viability and invasion capacity in vitro and in vivo

The aim of the present study was to determine the effect of AZD8055 on proliferation, apoptosis and glycolysis in the human cervical cancer cell line HeLa and to investigate the underlying mechanism(s) of action. HeLa human cervical cancer cells were treated with 10 nM AZD8055 for 24, 48 or 72 h. MTT was used to determine cell proliferation. Annexin V/propidium iodide staining was used to determine cell apoptosis analyzed by fluorescence-activated cell sorting (FACS). Glycolytic activity was determined by measuring the activity of the key enzyme lactate dehydrogenase (LDH) and lactate production. RNA and protein expression were examined by qRT-PCR and western blotting, respectively. Treatment with AZD8055 inhibited proliferation and glycolysis, and induced apoptosis in HeLa cells in a time-dependent manner. During the prolonged treatment with AZD8055, the phosphorylation of mammalian target of rapamycin (mTOR) C1 substrates p70S6K and phosphorylation of the mTORC2 substrate Akt were deregulated, suggesting that the activity of mTOR was downregulated. Furthermore, our study showed that the expression of miR-143 was upregulated in a time-dependent manner in HeLa cells treated with AZD8055. In summary, the present study reveals a novel antitumor mechanism of AZD8055 in HeLa human cervical cancer cells.

/pdf/the-mtor-inhibitor-azd8055-inhibits-proliferation-and-2xswjvggse.pdf

The mTOR inhibitor AZD8055 inhibits proliferation and glycolysis in cervical cancer cells

Predictors for local recurrence and distant metastasis of mucinous colorectal adenocarcinoma.

Objective To investigate the prognostic factors of young, middle-age and old-age colorectal cancer patients in order to improve the treatment in the future. Methods Colorectal cancer patients（ n=842） who had undergon curative resection were divided into three groups according their age： young group（≤40 years）, middle-age group（41 to 64 years ） and old group （≥65 years）. Thirty-five clinical factors in the three groups were analyzed and compared by univariate survival and multivariate analysis. Cox proportional hazards regression model was used with SPSS statistic software. Results The overall 5-, 10-and 15-year survival rates were 66.3%, 54.2% and 48.5% respectively. The 5-and 10-year survival rates were 53.0% and 42.7% in the young group, which were lower than those in the other two groups. Cox proportional hazards regression model demonstrated that Dukes stage and family history of cancer were common prognostic factors in both young and middle-age groups; chronic constipation was an independent prognostic factor in middle-age group; bowel obstruction, length of operating time and number of metastatic lymph nodes were prognostic factors in the older group. In the young group, the symptomatic duration was not demonstrated as a prognostic factor. The 5- and 10-year survival rates were 82.6% and 64.5% in Dukes A stage; 73.3% and 67.4% in B stage; 37.3% and 27.0% in C stage; 33.3% and 22.2% in D stage. The survival rates in Dukes A and B stages were similar, but in Dukes C and D stages they were lower than those of the middle-age and older groups if the patient had the same stage of disease. in the young colorectal cancer patients with family cancer history, the 5- and 10-year survival rates were 73.1% and 64.5 %, which were better than those of patients without it （48.1% and 37.3 % ）. Conclusion In young colorectal cancer patients, the survival rate is lower than those in the middle-age and old patients. Family cancer history and/or advanced Dukes stage are poor prognostic factors, whereas the symptomatic duration is not demonstrated as a poor prognostic factor. The prognostic factors affecting the survival after surgical treatment may be different in different age groups of colorectal cancer patients.

Key words: 
Colorectal neoplasms/surgery;  Colorectal neoplasms/pathology;  Prognosis;  Age

Multivariate analysis of prognostic factors in colorectal cancer patients with different ages

Objective To investigate the effect of microRNA143 on cell proliferation and K-ras expression in colorectal carcinoma. Methods Northern blot was used to examine the expression of miR-143 in colorectal carcinoma and adjacent normal tissues, h miR-143 expression vector was constructed and transfected into a human colon adenocarcinoma cell line SW480. Cell proliferation was evaluated by MTT assay. RT-PCR and Western blot were used to examine the expression of K-ras oncogene in transfected cells. Results The level of mature miR-143 was lower in tumors compared with adjacent normal tissues in 81% of colorectal carcinoma specimens. In transfected cells, the increased accumulation of miR-143 inhibited the cell proliferation, and resulted in approximately 40. 3% decrease of K-ras protein levels, but had no effect on level of K-ras mRNA. Conclusion The increased accumulation of miR-143 inhibits the proliferation of transfected cells, and results in down-regulation of K-ras protein in colorectal carcinoma.

Key words: 
MicroRNA143 ;  Colorectal neoplasms;  Cellular proliferation ;  K-ras

Effect of microRNA143 expression on cell proliferation in colonic carcinoma

Objective To analyze the adenomatous polyposis coli (APC)gene mutations in familial adenomatous polyposis(FAP)in Chinese.Methods DNA was extracted from blood samples taken from 31 FAP families ,and all formance liquid chromatography followed by sequencing if abnormal profile Was detected.Results Twelve categories ofAPC gene mutations were found in 15 FAP families(48.39%)including 4 novel mutations in coding region and 3 mutations in introns.The 4 novel mutations in coding region were frameshift mutations and located in codons 255,677,1192 and 1403 respectively.Most mutations were clustered in exon 15 of APC gene leading to frameshift and accounted for 86.67%.Others were nonsense mutatiom(13.33%).Conclusion The mutation rate of the APC gene in this group of Chinese FAP families was about 48.39%,and 4 novel mutations were detected.Frameshift mutation was the major mutation type in Chinese FAr and mainly located in exon 15.

Key words: 
familial adenomatous polyposis; denaturing high performance liquid chromatography; adenomatous polyposis coli gene; mutatio

Detection of adenomatous polyposis coli gene mutations in 31 familial adenomatous polyposis families by using denaturing high performance liquid chromatography

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Multivariate analysis of prognostic factors in colorectal cancer patients with different ages

Effect of microRNA143 expression on cell proliferation in colonic carcinoma

Detection of adenomatous polyposis coli gene mutations in 31 familial adenomatous polyposis families by using denaturing high performance liquid chromatography