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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

MiR-195 inhibits tumor growth and metastasis of osteosarcoma in vivo(A)I, Representative photographs of tumors on the right leg of mouse; II, Representative photographs of orthotopic tumors harvested 42 days after inoculation. III, Representative macroscopic pictures of mouse lungs, 42 days after inoculation. (B) Representative photographs of H&E stained spontaneous orthotopic tumors at a magnification of 400×. (C) Representative photographs of H&E stained spontaneous lung metastases at a magnification of 400×. (D) Tumor growth curves measured after the inoculation. The length (L) and width (W) of tumor measured every 7days after inoculation, and the volume of tumor was calculated according to the formula: volume = 1/2×L×W2. (E) Orthotopic tumor weights 42 days after inoculation. Data are presented as means±SD. (F) Graph displaying the total number of tumor nodules per lung in three groups. Data are presented as means±SD. (G) 42 days after inoculation, miR-195 expression levels in orthotopic tumors were tested and showed in relative miR-195 levels. 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 4: MiR-195 inhibits tumor growth and metastasis of osteosarcoma in vivo(A)I, Representative photographs of tumors on the right leg of mouse; II, Representative photographs of orthotopic tumors harvested 42 days after inoculation. III, Representative macroscopic pictures of mouse lungs, 42 days after inoculation. (B) Representative photographs of H&E stained spontaneous orthotopic tumors at a magnification of 400×. (C) Representative photographs of H&E stained spontaneous lung metastases at a magnification of 400×. (D) Tumor growth curves measured after the inoculation. The length (L) and width (W) of tumor measured every 7days after inoculation, and the volume of tumor was calculated according to the formula: volume = 1/2×L×W2. (E) Orthotopic tumor weights 42 days after inoculation. Data are presented as means±SD. (F) Graph displaying the total number of tumor nodules per lung in three groups. Data are presented as means±SD. (G) 42 days after inoculation, miR-195 expression levels in orthotopic tumors were tested and showed in relative miR-195 levels. 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: Given these findings in vitro, animal studies were conducted to evaluate the effect of miR-195 on orthotopic tumor growth and metastasis in athymic nude mice. Three groups of stable cells (OE, control and KD SOSP-9607) were injected into proximal tibia of young nude mice as described in methods, respectively. The results showed that cells formed progressively growing solid tumors in all mice. By contrast, cells in OE groups produced much smaller and slowly growth tumors than the KD groups (Fig. 4A, B, p<0.05). The mean tumor weight±SD of orthotopic tumors in OE group and KD group was 1.008±0.219 g and 2.104±0.187 g, respectively (Fig. 4A, B). Lung tissue of nude mice was also removed to observe the level of metastases. The result showed that the number of metastatic node was dramatically reduced in the nude mice in OE groups when compared to the other groups (Fig. 4A, B). Then tumor and metastases were confirmed based on histopathological evaluation (Fig. 4B, C). Meanwhile, the miR-195 expression levels in the orthotopic tumors were higher in the OE groups as compared with other groups (Fig. 4G). It indicates that exogenous miR-195 can significantly inhibit the tumor growth and metastasis of OS in vivo.


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)

MiR-195 inhibits tumor growth and metastasis of osteosarcoma in vivo(A)I, Representative photographs of tumors on the right leg of mouse; II, Representative photographs of orthotopic tumors harvested 42 days after inoculation. III, Representative macroscopic pictures of mouse lungs, 42 days after inoculation. (B) Representative photographs of H&E stained spontaneous orthotopic tumors at a magnification of 400×. (C) Representative photographs of H&E stained spontaneous lung metastases at a magnification of 400×. (D) Tumor growth curves measured after the inoculation. The length (L) and width (W) of tumor measured every 7days after inoculation, and the volume of tumor was calculated according to the formula: volume = 1/2×L×W2. (E) Orthotopic tumor weights 42 days after inoculation. Data are presented as means±SD. (F) Graph displaying the total number of tumor nodules per lung in three groups. Data are presented as means±SD. (G) 42 days after inoculation, miR-195 expression levels in orthotopic tumors were tested and showed in relative miR-195 levels. 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 4: MiR-195 inhibits tumor growth and metastasis of osteosarcoma in vivo(A)I, Representative photographs of tumors on the right leg of mouse; II, Representative photographs of orthotopic tumors harvested 42 days after inoculation. III, Representative macroscopic pictures of mouse lungs, 42 days after inoculation. (B) Representative photographs of H&E stained spontaneous orthotopic tumors at a magnification of 400×. (C) Representative photographs of H&E stained spontaneous lung metastases at a magnification of 400×. (D) Tumor growth curves measured after the inoculation. The length (L) and width (W) of tumor measured every 7days after inoculation, and the volume of tumor was calculated according to the formula: volume = 1/2×L×W2. (E) Orthotopic tumor weights 42 days after inoculation. Data are presented as means±SD. (F) Graph displaying the total number of tumor nodules per lung in three groups. Data are presented as means±SD. (G) 42 days after inoculation, miR-195 expression levels in orthotopic tumors were tested and showed in relative miR-195 levels. 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: Given these findings in vitro, animal studies were conducted to evaluate the effect of miR-195 on orthotopic tumor growth and metastasis in athymic nude mice. Three groups of stable cells (OE, control and KD SOSP-9607) were injected into proximal tibia of young nude mice as described in methods, respectively. The results showed that cells formed progressively growing solid tumors in all mice. By contrast, cells in OE groups produced much smaller and slowly growth tumors than the KD groups (Fig. 4A, B, p<0.05). The mean tumor weight±SD of orthotopic tumors in OE group and KD group was 1.008±0.219 g and 2.104±0.187 g, respectively (Fig. 4A, B). Lung tissue of nude mice was also removed to observe the level of metastases. The result showed that the number of metastatic node was dramatically reduced in the nude mice in OE groups when compared to the other groups (Fig. 4A, B). Then tumor and metastases were confirmed based on histopathological evaluation (Fig. 4B, C). Meanwhile, the miR-195 expression levels in the orthotopic tumors were higher in the OE groups as compared with other groups (Fig. 4G). It indicates that exogenous miR-195 can significantly inhibit the tumor growth and metastasis of OS in vivo.

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