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MicroRNA profiling identifies MiR-195 suppresses osteosarcoma cell metastasis by targeting CCND1.

Han K, Chen X, Bian N, Ma B, Yang T, Cai C, Fan Q, Zhou Y, Zhao TB - Oncotarget (2015)

Bottom Line: MicroRNAs are important regulators in tumorigenesis and tumor progression.One of the top ranked miRNAs-miR-195 was identified highly expressing in lowly metastatic cells.Results indicated lowly expressed miR-195 or highly CCND1 correlated with positive overall survival and their expression inverse relate to each other.

View Article: PubMed Central - PubMed

Affiliation: Department of Spinal Cord Injury, General Hospital of Jinan Military Area Command of Chinese PLA, Jinan, Shandong, People's Republic of China.

ABSTRACT
Metastasis is a leading cause of mortality for osteosarcoma patients. The molecular pathological mechanism remains to be elucidated. In the previously study, we established two osteosarcoma cell lines with different metastatic potentials. Differential expressed genes and proteins regarding metastatic ability have been identified. MicroRNAs are important regulators in tumorigenesis and tumor progression. In this study, microRNA microarray was used to assess the differential expressed miRNAs level between these two cell lines. One of the top ranked miRNAs-miR-195 was identified highly expressing in lowly metastatic cells. It was showed that over-expression of miR-195 substantially inhibits migration and invasion of osteosarcoma cells in vitro and pulmonary metastasis formation in vivo. Meanwhile, CCND1 was identified as the target gene of miR-195 and further studied. More importantly, Using real-time PCR, we evaluated the expression of miR-195 and CCND1 in osteosarcoma samples from 107 frozen biopsy tissues and 99 formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues. Results indicated lowly expressed miR-195 or highly CCND1 correlated with positive overall survival and their expression inverse relate to each other. In summary, our study suggests miR-195 function as a tumor metastasis suppressor gene by down-regulating CCND1 and can be used as a potential target in the treatment of osteosarcoma.

No MeSH data available.


Related in: MedlinePlus

Expression level of miR-195 in 107 OS frozen samples and 99 osteosarcoma formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues(A) Relative levels of miR-195 in 107 surgical specimens of osteosarcoma and matched adjacent noncancerous tissues(NAT) was quantified by real-time RT-PCR. Data were presented as log2 fold change (ΔΔCt values, Tumor/noncancerous tissues, T/N). (B) Means of miR-195 relative levels for 107 surgical specimens of osteosarcoma and the matched adjacent noncancerous tissues. Data were presented as 2−ΔΔCt values (p=0.291>0.05). (C) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.0012) than those with low miR-195 expression. The median miR-195 expression level (0.92479) in the tumor samples was chosen as the cut-off point. (D) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma FFPE patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.019) than those with low miR-195 expression. The median miR-195 expression level (5.0214) in the tumor samples was chosen as the cut-off point. The data were presented as the means ±SD, Columns, mean of four independent experiments; bars, SD; * P < 0.05, ** P < 0.01, *** P < 0.001.
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Figure 5: Expression level of miR-195 in 107 OS frozen samples and 99 osteosarcoma formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues(A) Relative levels of miR-195 in 107 surgical specimens of osteosarcoma and matched adjacent noncancerous tissues(NAT) was quantified by real-time RT-PCR. Data were presented as log2 fold change (ΔΔCt values, Tumor/noncancerous tissues, T/N). (B) Means of miR-195 relative levels for 107 surgical specimens of osteosarcoma and the matched adjacent noncancerous tissues. Data were presented as 2−ΔΔCt values (p=0.291>0.05). (C) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.0012) than those with low miR-195 expression. The median miR-195 expression level (0.92479) in the tumor samples was chosen as the cut-off point. (D) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma FFPE patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.019) than those with low miR-195 expression. The median miR-195 expression level (5.0214) in the tumor samples was chosen as the cut-off point. The data were presented as the means ±SD, Columns, mean of four independent experiments; bars, SD; * P < 0.05, ** P < 0.01, *** P < 0.001.

Mentions: To determine whether miR-195 expression correlates with clinical outcome in patients, we measured its levels in 107 pairs of OS frozen samples and corresponding noncancerous bone tissues by quantitative real time RT-PCR. Totally, miR-195 expression was decreased in 93 of 107 (86.92%) tumor samples compared with their non-malignant counterparts (Fig. 5A). The results showed that the relative level of miR-195 expression in OS tissues (mean± SD: 0.9248±0.4869) was significantly lower than that in matched nontumor adjacent tissues (NATs) (mean ± SD: 1.8762 ±0.6724; p<0.001) (Fig. 5B).


MicroRNA profiling identifies MiR-195 suppresses osteosarcoma cell metastasis by targeting CCND1.

Han K, Chen X, Bian N, Ma B, Yang T, Cai C, Fan Q, Zhou Y, Zhao TB - Oncotarget (2015)

Expression level of miR-195 in 107 OS frozen samples and 99 osteosarcoma formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues(A) Relative levels of miR-195 in 107 surgical specimens of osteosarcoma and matched adjacent noncancerous tissues(NAT) was quantified by real-time RT-PCR. Data were presented as log2 fold change (ΔΔCt values, Tumor/noncancerous tissues, T/N). (B) Means of miR-195 relative levels for 107 surgical specimens of osteosarcoma and the matched adjacent noncancerous tissues. Data were presented as 2−ΔΔCt values (p=0.291>0.05). (C) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.0012) than those with low miR-195 expression. The median miR-195 expression level (0.92479) in the tumor samples was chosen as the cut-off point. (D) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma FFPE patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.019) than those with low miR-195 expression. The median miR-195 expression level (5.0214) in the tumor samples was chosen as the cut-off point. The data were presented as the means ±SD, Columns, mean of four independent experiments; bars, SD; * P < 0.05, ** P < 0.01, *** P < 0.001.
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Figure 5: Expression level of miR-195 in 107 OS frozen samples and 99 osteosarcoma formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues(A) Relative levels of miR-195 in 107 surgical specimens of osteosarcoma and matched adjacent noncancerous tissues(NAT) was quantified by real-time RT-PCR. Data were presented as log2 fold change (ΔΔCt values, Tumor/noncancerous tissues, T/N). (B) Means of miR-195 relative levels for 107 surgical specimens of osteosarcoma and the matched adjacent noncancerous tissues. Data were presented as 2−ΔΔCt values (p=0.291>0.05). (C) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.0012) than those with low miR-195 expression. The median miR-195 expression level (0.92479) in the tumor samples was chosen as the cut-off point. (D) Decreased expression of miR-195 was correlated with poor survival in osteosarcoma FFPE patients. Log rank tests show that patients with high miR-195 expression survived statistically significantly longer (p = 0.019) than those with low miR-195 expression. The median miR-195 expression level (5.0214) in the tumor samples was chosen as the cut-off point. The data were presented as the means ±SD, Columns, mean of four independent experiments; bars, SD; * P < 0.05, ** P < 0.01, *** P < 0.001.
Mentions: To determine whether miR-195 expression correlates with clinical outcome in patients, we measured its levels in 107 pairs of OS frozen samples and corresponding noncancerous bone tissues by quantitative real time RT-PCR. Totally, miR-195 expression was decreased in 93 of 107 (86.92%) tumor samples compared with their non-malignant counterparts (Fig. 5A). The results showed that the relative level of miR-195 expression in OS tissues (mean± SD: 0.9248±0.4869) was significantly lower than that in matched nontumor adjacent tissues (NATs) (mean ± SD: 1.8762 ±0.6724; p<0.001) (Fig. 5B).

Bottom Line: MicroRNAs are important regulators in tumorigenesis and tumor progression.One of the top ranked miRNAs-miR-195 was identified highly expressing in lowly metastatic cells.Results indicated lowly expressed miR-195 or highly CCND1 correlated with positive overall survival and their expression inverse relate to each other.

View Article: PubMed Central - PubMed

Affiliation: Department of Spinal Cord Injury, General Hospital of Jinan Military Area Command of Chinese PLA, Jinan, Shandong, People's Republic of China.

ABSTRACT
Metastasis is a leading cause of mortality for osteosarcoma patients. The molecular pathological mechanism remains to be elucidated. In the previously study, we established two osteosarcoma cell lines with different metastatic potentials. Differential expressed genes and proteins regarding metastatic ability have been identified. MicroRNAs are important regulators in tumorigenesis and tumor progression. In this study, microRNA microarray was used to assess the differential expressed miRNAs level between these two cell lines. One of the top ranked miRNAs-miR-195 was identified highly expressing in lowly metastatic cells. It was showed that over-expression of miR-195 substantially inhibits migration and invasion of osteosarcoma cells in vitro and pulmonary metastasis formation in vivo. Meanwhile, CCND1 was identified as the target gene of miR-195 and further studied. More importantly, Using real-time PCR, we evaluated the expression of miR-195 and CCND1 in osteosarcoma samples from 107 frozen biopsy tissues and 99 formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues. Results indicated lowly expressed miR-195 or highly CCND1 correlated with positive overall survival and their expression inverse relate to each other. In summary, our study suggests miR-195 function as a tumor metastasis suppressor gene by down-regulating CCND1 and can be used as a potential target in the treatment of osteosarcoma.

No MeSH data available.


Related in: MedlinePlus