1.The effect of siRNA targeting MIF on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice
Yamin WANG ; Lijing WANG ; Rongjiao YANG ; Jieyi CAI ; Lihao WU ; Xingxiang HE
Chinese Journal of General Surgery 2011;26(5):376-380
Objective To analyze the effect of siRNA targeting MIF( MIFsiRNA) on the growth of colorectal cancer xenografts and the life quality of tumor-bearing mice.Methods BALB/C mouse model carring colorectal cancer was established.Thirty mice were divided into three groups randomly and managed respectively with intratumor injection of DEPC water, MIFsiRNA(0.15 nmol/g) and non-specific siRNA (0.15 nmol/g), respectively twice a week for consecutively 4 weeks.Drinking water, fodder consumed and body weight was recorded daily, and tumor volume was measured once a week.Mice were sacrificed after four weeks.ELISA and immunohistochemistry were used to detect the expression of MIF in serum and in tumor tissues.Spectrophotometric detection was used to detect caspase-3 protein.TUNEL was used to detect apoptotic cells.Results MIF expression in serum in MIFsiRNA group was lower than the other two groups [(22 ± 6) ng/ml vs (32 ± 8) ng/ml and (33 ± 8) ng/ml, P < 0.01]; MIF expression in tissues was less than the other two groups [(85 ± 20) /500 vs.(423 ± 23) /500 and (442 ± 31) /500, P < 0.01]; Tumor was smaller than the other two groups at third and fourth week (P < 0.01) ; Tumor weight was significantly less than the other two groups [(1.93 ±0.21) g vs (4.40 ±0.30) g and (5.25 ±0.44) g, P<0.01]; Mice in MIFsiRNA group were healthier than the other two groups as judged by water and fodder consumption (P < 0.01 ) , while weight change was not significantly different among the three groups ( P > 0.05 ).Caspase-3 protein in tissues was higher than the other two groups [(0.74 ±0.06) μg vs (0.57 ±0.08) μg and (0.56 ±0.02) μg, P <0.01]; Apoptosis cells in tissues were higher than the other two groups [(12 ± 2)/ 100 个vs 0 and 0, P < 0.01].Conclusions Knockdowning MIF gene expression inhibits the growth of colorectal cancer xenografts and improves life quality of tumor-bearing mice, possibly by a mechanism in which MIFsiRNA activates caspase-3 promoting cell apoptosis.
2.Role of miR-24 in the regulation of 3T3-L1 adipicyte differentiation and the expression of fatty acid binding protein
Limei YAN ; Qiuyan LIANG ; Jinhua XU ; Jianyong WU ; Rongjiao WANG ; Xiaohua YU ; Jingou XU ; Hesheng OU
Chinese Pharmacological Bulletin 2010;26(4):483-487
Aim To explore the effect of microRNAs on the differentiation of 3T3-L1 adipocytes and the expression of adipo-related gene-fatty acid binding protein during the adipocyte differentiation.Methods adipo-related microRNAs during 3T3-L1 adipocyte differentiation were screened and identified by micorRNA microarray.Constructed high-expression plasmids of the adipo-related microRNAs,were transfected into the 3T3-L1 preadipocytes by lipofectamine.While the effect of adipo-related microRNAs on the course of 3T3-L1 adipocyte differentiation was observed,the protein and mRNA expression level of fatty acid binding protein(FABP4)were analyzed by Western blot and RT-PCR during 3T3-L1 adipocyte differentiation.Results The expression profiles of microRNAs have significant changed during 3T3-L1 adipocyte differentiation,in which 35 microRNAs among them down-relation,the most lowly expression is miR-24;17 microRNAs among them up-relation,the most highly expression is miR-21.MiR-24 significantly inhibited adipocyte differentiation and maturity,while miR-21 have no significant effect.MiR-24 significantly inhibited the expression of FABP4,but had no effect on the level of its mRNA;miR-21 had no effect on the expression of protein and mRNA of FABP4.Conclusion There exist adipogenic-related microRNAs during 3T3-L1 adipocyte differentiation; miR-24 play an important role in the regulation of 3T3-L1 preadipocyte differentiation into adipocyte and the(FABP4)protein expression.
3.Expression of macrophage migration inhibitory factor in peripheral blood of patients with Hashimoto's thyroiditis and its clinical significance
Haibo XUE ; Lei MA ; Xiuyun WANG ; Junying LIU ; Rongjiao ZHOU ; Li ZHANG
Chinese Journal of Endocrinology and Metabolism 2012;28(9):741-743
The levels of macrophage migration inhibitory factor (MIF) in peripheral blood mononuclear cell (PBMC) and serum in patients with Hashimoto's thyroiditis (HT) were detected by real-time PCR and ELISA,respectively.The results revealed that the expression of MIF mRNA in PBMC ( Z =-4.276,P<0.01 ) and protein level in serum ( Z=-5.358,P<0.01 ) were increased in HT patients,and positively correlated with thyroid specific autoantibodies and TSH levels.