中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
3期
290-291
,共2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
膀胱癌%Fibulin-5%增殖
膀胱癌%Fibulin-5%增殖
방광암%Fibulin-5%증식
Urothelial carcinoma%Fibulin-5%Proliferation
目的 观察Fibulin-5对膀胱癌细胞增殖的影响.方法 采用脂质体介导法将构建好的p-EGFP-N1-Fibulin-5表达载体转染到膀胱癌5637细胞中.通过噻唑蓝(MTT)比色法和细胞计数法检测Fibulin-5表达上调后膀胱癌细胞生长率的变化.结果 酶切鉴定及DNA测序结果显示Fibulin-5 cDNA插入正确;转染48 h后,观测到p-EGFP-N1-F5表达载体转染成功的细胞均有不同程度的绿色荧光表达.培养第7天,MTT法测得转染Fibulin-5、转染空载体和未转染的膀胱癌细胞的吸光值分别为0.330±0.025、0.640 ±0.030和0.72±0.060.绘制生长曲线发现转染Fibulin-5的膀胱癌细胞的生长率明显低于转染空载体(P＜0.05)和未转染的细胞(P＜0.05).细胞计数结果和MTT一致.结论 Fibulin-5可能通过抑制膀胱癌细胞的增殖而在膀胱癌中起抑癌基因的作用.
目的 觀察Fibulin-5對膀胱癌細胞增殖的影響.方法 採用脂質體介導法將構建好的p-EGFP-N1-Fibulin-5錶達載體轉染到膀胱癌5637細胞中.通過噻唑藍(MTT)比色法和細胞計數法檢測Fibulin-5錶達上調後膀胱癌細胞生長率的變化.結果 酶切鑒定及DNA測序結果顯示Fibulin-5 cDNA插入正確;轉染48 h後,觀測到p-EGFP-N1-F5錶達載體轉染成功的細胞均有不同程度的綠色熒光錶達.培養第7天,MTT法測得轉染Fibulin-5、轉染空載體和未轉染的膀胱癌細胞的吸光值分彆為0.330±0.025、0.640 ±0.030和0.72±0.060.繪製生長麯線髮現轉染Fibulin-5的膀胱癌細胞的生長率明顯低于轉染空載體(P＜0.05)和未轉染的細胞(P＜0.05).細胞計數結果和MTT一緻.結論 Fibulin-5可能通過抑製膀胱癌細胞的增殖而在膀胱癌中起抑癌基因的作用.
목적 관찰Fibulin-5대방광암세포증식적영향.방법 채용지질체개도법장구건호적p-EGFP-N1-Fibulin-5표체재체전염도방광암5637세포중.통과새서람(MTT)비색법화세포계수법검측Fibulin-5표체상조후방광암세포생장솔적변화.결과 매절감정급DNA측서결과현시Fibulin-5 cDNA삽입정학;전염48 h후,관측도p-EGFP-N1-F5표체재체전염성공적세포균유불동정도적록색형광표체.배양제7천,MTT법측득전염Fibulin-5、전염공재체화미전염적방광암세포적흡광치분별위0.330±0.025、0.640 ±0.030화0.72±0.060.회제생장곡선발현전염Fibulin-5적방광암세포적생장솔명현저우전염공재체(P＜0.05)화미전염적세포(P＜0.05).세포계수결과화MTT일치.결론 Fibulin-5가능통과억제방광암세포적증식이재방광암중기억암기인적작용.
Objective To study the effect of Fibulin-5 on the proliferation of bladder cancer cells.Methods Fibulin-5 cDNA was amplified by polymerase chain reaction(PCR)and cloned into pMD.19T simple vectors.The pMD-19T-Fibulin-5 vector was digested by restriction endonueleases Xhol and EcoRI to generate a-1.5 kb XhoI-Fibulin5-EcoRI fragment that was then ligated into the identical sites in a p-EGFP N1 plasmid to synthesize the p-EGFP-Fibulin-5 plasmid.Next,the p-EGFP-Fibulin-5 plasmid was transfected into bladder cancer cell line 5637.Finally,the proliferation of transfected cells was measured by MMT assay and cell number count.Results The results of DNA sequencing and restriction endonucleases digestion suggested the correct construction of eukaryon expression vector p-EGFP-Fibulin-5.At the 48th h after transfection,fluorescent microscopy(488 nm)revealed that the transfected cells emitted green fluorescence.The absorbance of Fibulin-5-transfected cells by MTT assay Was 0.330±0.025 in the 7th day of eell culture,which was significantly lower than that of untransfected cells(0.640±0.030)or empty vector-transfected cells (0.720±0.060)(P＜0.05).The result of cell number count Was similar to that of MTT assay.Conclusion Fibulin-5 might act as a tumor suppressor gene of bladder cancer by inhibiting the proliferation of bladder cancer cells.
