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The Study on Reversing Multidrug Resistance of Human Bladder Carcinoma Drug-resistant Cell Line BIU-87/ADM by Tetrandrine

Author: LiuJinYu
Tutor: LiYongSheng
School: Fujian Medical
Course: Surgery
Keywords: Tetrandrine Adriamycin Bladder cancer Resistant Apoptosis resistance
CLC: R285.5
Type: Master's thesis
Year: 2011
Downloads: 22
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Abstract


Objective: 1. To explore Tetrandrine (tetrandrine, TET) to reverse the effect and mechanism of human bladder cancer resistant strains BIU-87/ADM multidrug resistance (multidrug resisitance, MDR). 2 confirmed tetrandrine can reverse the resistance of human bladder cancer of resistant strains BIU-87/ADM the apoptosis, and to explore its possible mechanism. Method: 1. Application of CCK law screening out the BIU-87 and no significant cytotoxicity BIU-87/ADM cells tetrandrine concentration; the CCK assay tetrandrine reverse multidrug resistance activity; reverse transcription - polymerase chain reaction (RT-PCR) and immunofluorescence tetrandrine of MDR1mRNA and P-gp protein expression were detected; the TUNEL assay tetrandrine BIU-87 cell apoptosis and BIU-87/ADM. Annexin-V and PI double staining flow cytometry analysis tetrandrine of BIU-87 and BIU-87/ADM apoptosis. The method of immunocytochemistry and RT-PCR assay tetrandrine alkali the the survivin was BIU-87/ADM cells, of livin, caspase-3 expression levels. Results: 1 1.0μg/mL powder tetrandrine on BIU-87, and BIU-87/ADM cells had no significant cytotoxic effect (inhibition rate 10%); 1.0μg/mL tetrandrine significantly increased doxorubicin (adriamycin, ADM) on the resistant strains BIU-87/ADM cytotoxicity (P lt; 0.01), reversion index (reversal fold, RF) was 5.33, while the sensitive line BIU-87 No significant role (P gt; 0.05), RF is only 1.33; the 1.0μg/mL tetrandrine can significantly inhibit MDR1mRNA and P-gp protein expression in BIU-87/ADM; 1.0μg/mL tetrandrine can significantly enhance adriamycin-induced BIU-87 / ADM cell apoptosis (P lt; 0.01), BIU-87 cells (P gt; 0.05). BIU-87 cells, and BIU-87/ADM cell apoptosis rate. In 1.5μg/mL doxorubicin were (57.79 ± 4.73)% and (12.42 ± 3.24)%, illustrates BIU-87/ADM cell apoptosis resistance. The 1.0μg/mL tetrandrine alkali and 1.5μg/mL of doxorubicin role of co-48h, make BIU-87/ADM apoptosis rate increased to (40.86 ± 4.35)%, compared to the separate application doxorubicin differences statistics significance (P lt; 0.01). BIU-87 cell apoptosis rate (P gt; 0.05), and this tetrandrine apoptosis resistance reversal BIU-87/ADM; 1.0μg/mL tetrandrine and 1.5μg / mL doxorubicin BIU-87/ADM cell lines can be of survivin of livin expression levels significantly lower caspase-3 expression levels were significantly increased compared with doxorubicin difference was statistically significant (P lt; 0.01). Conclusion: The low concentrations of tetrandrine reversed BIU-87/ADM cell multidrug resistance, this role may tetrandrine inhibition of P-gp expression and induction chemotherapy drugs to promote apoptosis of bladder cancer . Low concentrations of tetrandrine can reverse the BIU-87/ADM apoptosis resistance, this role of survivin may tetrandrine and doxorubicin combined with down of livin expression and upregulation of caspase-3 expression related.

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