Effect of chemokine stromal cell-derived factor-1 and its receptor CXCR4 on invasion capacity and intralu-minal implantation of human bladder cancer cells
- VernacularTitle:基质细胞衍生因子-1及其受体 CXCR4对膀胱癌细胞侵袭能力及腔内种植的影响
- Author:
Delin YANG
;
Qian HUO
;
Yishui WANG
;
Xusheng YANG
;
Kai WANG
;
Jiansong WANG
;
Hongyi XU
;
Haifeng WANG
- Publication Type:Journal Article
- Keywords:
CXCR4;
ShRNA;
Bladder cancer;
Invasion
- From:
Journal of Medical Postgraduates
2014;(10):1028-1032
- CountryChina
- Language:Chinese
-
Abstract:
Objective Bladder cancer , which has a high rate of recurrence and invasion , is the most common genitourinary cancer.The article was to study the effect of specific chemokine receptor CXCR 4 on invasion capacity and intraluminal implantation of human bladder cancer cells . Methods A CXCR4 specific recombinant plasmid vector (short hairpin, shRNA) was constructed to select those cells which could inhibit the expression of CXCR 4, and these cells were divided into blank control group , negative control plasmid group and recombinant plasmid group (pshRNA-CXCR4-1, pshRNA-CXCR4-2).RT-PCR and immunofluorescence technique were used to detect the mRNA and protein expression of CXCR 4 respectively .Invasion capability in vitro of the cells was evaluated by Boyden chamber .20 nude mice were randomly divided into experimental group and control group ( n=10 ) .The experimental group was established by injection of 100μL shRNA-EJ-M3 into the bladder , while the control group was established by injection of 100μL EJ-M3, aiming to detect the effect of shRNA-CXCR4 on intraluminal implantation of human bladder cancer cells . Results The CXCR4 mRNA expression of the pshRNA-CXCR4-1 group (62.05 ± 1.35) was significantly lower than that of blank control group (174.38 ±1.96, P <0.05 ) and negative control plasmid group (166.27 ±1.82, P <0.05).There was no significant difference among the CXCR4 mRNA expression of the pshRNA-CXCR4-2 group, blank control group and negative control plasmid group ( 182.58 ± 4.2, 174.38 ±1.96, 166.27 ±1.82, P>0.05).In immunofluores-cence experiment, the red cell amount of the pshRNA-CXCR4-1 group(32.24 ±2.23) was lower than that of the blank control group (89.61 ±4.47,P<0.05) and negative control plasmid group (92.45 ±3.68, P<0.05).There was no significant difference among the red cell amount of the pshRNA-CXCR4-2 group, the blank control group and the negative control plasmid group (76.87 ±5.11, 89.61 ±4.47, 92.45 ±3.68, P>0.05).The Boyden chamber experiment showed that the number of penetrating cells of the pshRNA -CXCR4-1 group (39.67 ±8.45) was significantly lower than that of the blank control group (135.33 ±9.28, P<0.05) and that of the negative control plasmid group(123.63 ±6.36, P<0.05).As to the intraluminal implanting capability, the difference between the ex-perimental group and the control group of statistical significance (10%vs 70%,P<0.01). Conclusion CXCR4 shRNA can inhibit the expression of CXCR4 and significantly decrease the invasion capacity and intraluminal implantation of human bladder cancer cells .
