[The effect of PPS on location of rBCG in bladder cancer cell line T24].
OBJECTIVE
To investigate the effect of polyporus polysaccharide (PPS) on location of green fluorescent recombinant protein BCG in human bladder cancer cell line T24.


METHODS
A green fluorescent protein expressed with rBCG was used as reporter gene, and laser scanning confocal microscopy and flow cytometry(FCM) analysis were performed to examine the alteration of T24 cells stimulated with rBCG or rBCG plus PPS.


RESULTS
Green fluorescence (86.335+/-5.856) was observed obviously in the cytoplasm of T24 cells after interaction with rBCG for 24 h. By FCM we found scatter dot signal in SS and FS channel from rBCG groups was stronger that a control at 24 h and 48 h respectively. Meanwhile the population of T24 cells deflected to upper right and the percentage of GFP(+) T24 cells detected in FL1 channel was (8.7+/-1.572)%, (13.8+/-2.31)% (P<0.05 compared to the control) respectively. For rBCG plus PPS groups, the fluorescence intensity increased in the nucleus (72.603+/-1.165), while decreased in the cytoplasm (93.06+/-0.958), thus the nucleus/plasm ratio increased (0.78+/-0.005). Compared to rBCG group (62.832+/-2.909, 105.306+/-6.393, 0.597+/-0.012), the difference was statistically significant (P<0.05).


CONCLUSIONS
rBCG is able to enter directly into the cytoplasm of T24 cells.
Zeng
Xing
X
Central Laboratory, Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
Yang
Ming
M
Wang
Jia
J
Zhang
Xian
X
Sun
Jing-Bo
JB
Duan
Xiao-Jun
XJ
D001692:Biological Transport / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D004274:DNA, Recombinant; D005434:Flow Cytometry; D006801:Humans; D018613:Microscopy, Confocal; D009163:Mycobacterium bovis / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000494:pharmacology*; D001749:Urinary Bladder Neoplasms / Q000382:microbiology* / Q000473:pathology
D004274:DNA, Recombinant; D011134:Polysaccharides