中华泌尿外科杂志
中華泌尿外科雜誌
중화비뇨외과잡지
CHINESE JOURNAL OF UROLOGY
2010年
3期
190-193
,共4页
闫永吉%胡争%艾青%吴准%李俊%马鑫%李宏召%史涛坪%王保军%龚道静%倪栋
閆永吉%鬍爭%艾青%吳準%李俊%馬鑫%李宏召%史濤坪%王保軍%龔道靜%倪棟
염영길%호쟁%애청%오준%리준%마흠%리굉소%사도평%왕보군%공도정%예동
膀胱肿瘤%肿瘤侵润%Fibulin-5
膀胱腫瘤%腫瘤侵潤%Fibulin-5
방광종류%종류침윤%Fibulin-5
Bladder neoplasms%Neoplasm invasiveness%Fibulin-5
目的 探讨fibulin-5表达在膀胱尿路上皮癌中的影响作用. 方法 经尿道切除获取膀胱尿路上皮癌标本20例,其中低级别(G_1和G_2)13例、高级别(G_3)7例.蛋白质印迹法检测fibulin-5在膀胱尿路上皮癌和正常膀胱黏膜组织中的表达.将PCR扩增的fibulin-5 cDNA克隆到pMD-19T载体中,构建p-EGFP-fibulin-5质粒.脂质体介导法将p-EGFP-fibulin-5质粒转染到人膀胱癌5637细胞株内,流式细胞仪分选,经G418筛选形成稳定的表达克隆.Boyden小室法检测未转染、转染空载体和转染fibulin-5的膀胱癌细胞迁移和侵袭能力. 结果 正常膀胱黏膜、低级别和高级别膀胱尿路上皮癌组织中fibulin-5蛋白相对表达量分别为1.16±0.28、0.57±0.32和0.44±0.42.与正常膀胱黏膜相比,膀胱癌组织中fibulin-5蛋白表达显著下调(P<0.01),低级别和高级别癌之间差异无统计学意义(P>0.05).转染成功的膀胱癌5637细胞显示绿色荧光.未转染、转染空载体和转染fibulin-5的膀胱癌细胞的迁移细胞数分别为136.9±5.7、139.3±7.7和127.6±3.1,侵
袭细胞数分别为31.7±4.7、31.5±4.8和8.05±3.1.转染fibulin-5后膀胱癌细胞侵袭力明显降低
(P<0.01),其迁移力虽低于对照组但差异无统计学意义(P>0.05). 结论 Fibulin-5蛋白表达减
少可能是膀胱癌发生和发展的机制之一,过表达fibulin-5可以抑制膀胱癌细胞侵袭力.
目的 探討fibulin-5錶達在膀胱尿路上皮癌中的影響作用. 方法 經尿道切除穫取膀胱尿路上皮癌標本20例,其中低級彆(G_1和G_2)13例、高級彆(G_3)7例.蛋白質印跡法檢測fibulin-5在膀胱尿路上皮癌和正常膀胱黏膜組織中的錶達.將PCR擴增的fibulin-5 cDNA剋隆到pMD-19T載體中,構建p-EGFP-fibulin-5質粒.脂質體介導法將p-EGFP-fibulin-5質粒轉染到人膀胱癌5637細胞株內,流式細胞儀分選,經G418篩選形成穩定的錶達剋隆.Boyden小室法檢測未轉染、轉染空載體和轉染fibulin-5的膀胱癌細胞遷移和侵襲能力. 結果 正常膀胱黏膜、低級彆和高級彆膀胱尿路上皮癌組織中fibulin-5蛋白相對錶達量分彆為1.16±0.28、0.57±0.32和0.44±0.42.與正常膀胱黏膜相比,膀胱癌組織中fibulin-5蛋白錶達顯著下調(P<0.01),低級彆和高級彆癌之間差異無統計學意義(P>0.05).轉染成功的膀胱癌5637細胞顯示綠色熒光.未轉染、轉染空載體和轉染fibulin-5的膀胱癌細胞的遷移細胞數分彆為136.9±5.7、139.3±7.7和127.6±3.1,侵
襲細胞數分彆為31.7±4.7、31.5±4.8和8.05±3.1.轉染fibulin-5後膀胱癌細胞侵襲力明顯降低
(P<0.01),其遷移力雖低于對照組但差異無統計學意義(P>0.05). 結論 Fibulin-5蛋白錶達減
少可能是膀胱癌髮生和髮展的機製之一,過錶達fibulin-5可以抑製膀胱癌細胞侵襲力.
목적 탐토fibulin-5표체재방광뇨로상피암중적영향작용. 방법 경뇨도절제획취방광뇨로상피암표본20례,기중저급별(G_1화G_2)13례、고급별(G_3)7례.단백질인적법검측fibulin-5재방광뇨로상피암화정상방광점막조직중적표체.장PCR확증적fibulin-5 cDNA극륭도pMD-19T재체중,구건p-EGFP-fibulin-5질립.지질체개도법장p-EGFP-fibulin-5질립전염도인방광암5637세포주내,류식세포의분선,경G418사선형성은정적표체극륭.Boyden소실법검측미전염、전염공재체화전염fibulin-5적방광암세포천이화침습능력. 결과 정상방광점막、저급별화고급별방광뇨로상피암조직중fibulin-5단백상대표체량분별위1.16±0.28、0.57±0.32화0.44±0.42.여정상방광점막상비,방광암조직중fibulin-5단백표체현저하조(P<0.01),저급별화고급별암지간차이무통계학의의(P>0.05).전염성공적방광암5637세포현시록색형광.미전염、전염공재체화전염fibulin-5적방광암세포적천이세포수분별위136.9±5.7、139.3±7.7화127.6±3.1,침
습세포수분별위31.7±4.7、31.5±4.8화8.05±3.1.전염fibulin-5후방광암세포침습력명현강저
(P<0.01),기천이력수저우대조조단차이무통계학의의(P>0.05). 결론 Fibulin-5단백표체감
소가능시방광암발생화발전적궤제지일,과표체fibulin-5가이억제방광암세포침습력.
Objective To study the role of fibulin-5 in urothelial carcinoma of bladder. Methods Fibulin-5 expression was detected in bladder cancer tissues (13 cases of G_1 and G_2, 7 cases of G_3) and normal bladder mucosa samples by Western blotting assay. Fibulin-5 cDNA was amplified by PCR and cloned into pMD-19T simple vector. The pMD-19T-Fibulin-5 vector was digested by restriction endonucleases XhoI and EcoRI to generate a XhoI-Fibulin5-EcoRI fragment that was then ligated into the identical sites in p-EGFP-Nl plasmid to synthesize p-EGFP-Fibulin-5 plasmid. The p-EGFP-Fibulin-5 plasmid was finally transfected into bladder cancer cell line 5637. The migration and invasion of untransfected, vector-transfected and fibulin-5-transfected bladder cancer cells were measured by Boyden chamber assay. Results Compared to 1. 16 ±0. 28 in the normal control, the expression of fibulin-5 protein in low grade and high grade tumors were 0. 57±0. 32 and 0. 44±0. 42(P<0. 01, respectively). However, the difference between low grade and high grade tumors was not statistically significant (P>0. 05). The successfully transfected bladder cancer cells demonstrated green fluorescent light. The migrated cell number of fibulin-5-transfected cells was 127. 6 ± 3. 1 compared with 139. 3±7. 7 for vector-transfected cells and 136. 9±5. 7 for untransfected cells (P>0. 05, respectively). In contrast, the invaded cell number of fibulin-5-transfected cells was 8. 0±3. 1 compared with 31. 5±4. 8 for vector-transfected cells and 31. 7±4. 7 for untransfected cells (P<0. 01, respectively). Conclusion Fibulin-5 is down-regulated in urothelial carcinoma of bladder and acts as a tumor suppressor gene by inhibiting the invasion of bladder cancer cells.
