膀胱癌细胞在尿液中适应性生存的研究

The Adaptability of Bladder Cancer Cells in Urine

目的观察膀胱癌细胞是否能在尿液中存活及其存活时间.方法过滤除菌正常人中段晨尿,并行尿RT及生化检测.倒置显微镜观察在除菌尿液中培养的膀胱癌细胞.MTT法对比人膀胱癌细胞EJ、人肝癌细胞HepG2和人血管内皮细胞Ecv304分别在尿液、生理盐水和无血清1640中的存活情况.通过逐渐递加培养基中除菌晨尿,筛选高尿液适应性的膀胱癌细胞.结果除菌前后尿RT及生化无改变.EJ细胞在尿中12h贴壁,24h仍可见到存活细胞.经多元方差分析,在尿液中培养相同时间时,EJ较HepG2(P<0.001)和Ecv304(P=0.014)的存活数目多;HepG2与Ecv304的差异无统计学意义(P=0.062).高尿液适应性的膀胱癌细胞在尿液中,24h时有大量细胞存活,48h仍有细胞存活.结论膀胱癌细胞可以在尿液中存活,并且在尿液培养相同时间时较HepG2和Ecv304存活数目多.经尿液诱导后筛选出的膀胱癌细胞能在尿液中存活更长时间.

Objective To investigate the survival possibility and the survival time of bladder cancer cell in urine. Methods Urina sanguinis of healthy adult were degermed by filtration, and the urine before and aflerfihration were detected by RT and biochemistry. Bladder cancer cells EJ were cultured in filtrated urine, and the cell＇s survival and growth were observed by inverted microscope. E J, HepG2 and Eev304 were cultured in urine, isotonic Na chloride and basal culture in 96 orifices, and the OD value of cells of different times were detected by MTT. By gradual addition the urine into medium, the cells that had high adaptability in urine were selected and cultured. Results The RT and biochemistry of urine didn＇t change before and after filtration. The cells that were cultured in urine adhered on culture flask at about 12 h, and a few cells survived at 24 h. The OD value of cells of different times were detected by MTT, and muhifaetor variance analysis authentication showed that when the culture time was identical, the survival number of different cells were different （P = 0.001） , and Q analysis showed that the survival number of EJ were greater than HepG2 （P 〈 0.001） and Ecv304 （P = 0.014） , and the survival numbers of HepG2 and Eev304 had no significant difference （P = 0.062）. A large number of cells with high adaptability in urine survived in urine for 24 h, and there were survival cells when cultured for 48 h. Conclusions Bladder cancer cells can survive in urine, and the number of survival cells is greater than HepG2 and Ecv304 when the culture time is identical. The bladder cancer cells which were selected by gradually addition the urine to medium can survive longer in urine than before.