Effect of small interfering RNA on expression of β2M in pre-differentiated bone marrow mesenchymal stem cells |
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Authors: | DAI Bing FAN Shi-yang CHEN Long JIN Hai-dong CAI Jian-wu PAN Jun |
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Affiliation: | Department of Orthopedics, Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, China. |
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Abstract: | AIM:To study the effect of small interfering RNA (siRNA) on the expression of beta 2-microglo-bulin (β2M) in pre-differentiated bone marrow mesenchymal stem cells (BMSCs). METHODS:The β2M siRNA was transfected into the pre-differentiated BMSCs with Lipofectamine 2000. BMSCs were divided into transfection group, blank control group and negative control group. The expression of β2M at mRNA and protein levels was determined by real-time qPCR, Western blotting and laser confocal microscopy. The productions of aggrecan and type II collagen in pre-differentiated BMSCs were determined by toluidine blue staining and type Ⅱ collagen immunofluorescence. RESULTS:The results of real-time qPCR, Western blotting and laser confocal microscopy showed that siRNA successfully inhibited the expression of β2M at mRNA and protein levels in the pre-differentiated BMSCs. The results of toluidine blue and type Ⅱ collagen immunofluorescence staining showed that siRNA does not affect the productions of aggrecan and type Ⅱ collagen in the pre-differentiated BMSCs. CONCLUSION:siRNA targeting β2M reduces the expression of β2M in the pre-differentiated BMSCs and does not affect the chondrocyte characteristics of pre-differentiated BMSCs. |
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Keywords: | Small interfering RNA Beta 2-microglobulin Bone marrow mesenchymal stem cells |
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