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Objective To study the relationship between chemotherapeutic resistance to 5-fluorouracil (5-FU) and Golgi phosphoprotein 3 (GOLPH3) expression in human colon cancer cell line HT29.Methods HT29 cells were divided into four groups:control group,siRNA transfection group,experimental group 1 (30 μmol/L 5-FU),and experimental group 2 (siRNA-GOLPH3 + 30 μmol/L 5-FU).The silencing effect of GOLPH3 gene was detected by RT-PCR and Western blot.The tumor proliferation and formation ability of HT29 cell were measured respectively by MTT and plate colony formation assay.The protein expressions of GOLPH3,P-gp and β-catenin in HT29 cell were detected by Western blot.Results Compared with the control group,the expression level of GOLPH3 mRNA (1.000 ± 0.078 vs.0.147 ± 0.021,t =12.296,P ＜ 0.01) and protein(1.003 ± 0.235 vs.0.077 ± 0.399,t =6.723,P ＜ 0.01),the absorbance(A value) (1.000 ± 0.082 vs.0.769 ± 0.086,t =3.885,P ＜ 0.01) and the tumorigenesis (430 ±36 vs.297 ± 21,t =5.492,P ＜ 0.01) in transfection group significantly reduced.The A value (0.803 ± 0.101 vs.1.050 ± 0.140,t =2.855,P ＜ 0.05;0.242 ± 0.091 vs.1.050 ± 0.140,t =9.664,P ＜ 0.001) and the tumorigenesis (73 ± 8 vs.427 ± 29,t =20.363,P ＜ 0.001;305 ± 22 vs.427 ± 29,t =5.840,P ＜ 0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group,while the A value (t =8.264,P ＜ 0.001) and the tumorigenesis (t =17.346,P ＜ 0.001) of experimental group 2 significantly decreased than that experimental group 1.Compared to the control group (0.967 ± 0.094,1.001 ± 0.199,1.000 ± 0.101),the expressions of GOLPH3,P-gp and β-catenin in experimental group 1 (3.634 ± 0.574,2.424 ± 0.261,2.324 ± 0.418) significantly upregulated (t =7.944,P ＜ 0.01;t =7.520,P ＜ 0.01;t =5.330,P ＜ 0.01),while the expressions of these proteins in experimental group 2(0.520 ±0.176,0.466 ±0.159,0.933 ±0.049) were significantly lower than that in experimental group 1 (t =8.983,P ＜ 0.01;t =11.106,P ＜ 0.001;t =5.724,P ＜ 0.01).Conclusion Expression of GOLPH3 is associated with resistance to 5-FU in colon cancer HT29 cell by activating Wnt/β-catenin signal pathway.
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AIM:To investigate the effects of 17β-estradiol on the expression of macrophage migration inhibi-tory factor ( MIF) in cultured endometrial stromal cells from endometriosis .METHODS:Immunohistochemistry was used to identify the endometrial stromal cells .The expression of MIF at mRNA and protein levels was assessed by RT-PCR and Western blotting .RESULTS:Elevated expression of MIF at mRNA and protein levels was observed in the cultured endom-etrial stromal cells treated with 17β-estradiol.In endometrial stromal cells from the women with endometriosis , the level of MIF up-regulation by 17β-estradiol was significantly higher than that in the cells from the women without endometriosis . CONCLUSION:Endometrial stromal cells from endometriosis are more sensitive to 17β-estradiol, which up-regulates the expression of MIF and contribute to the pathogenesis and progression of endometriosis .