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Effect of Paeoniflorin on Drug Resistance of Human Gastric Cancer SGC7901/VCR Cells and Its Mechanism

Author: FangShenCun
Tutor: LiuPing
School: Nanjing Medical University
Course: Oncology
Keywords: Paeoniflorin Gastric cancer Apoptosis NF-κB Bcl-xl Bcl-2 P-gp
CLC: R735.2
Type: Master's thesis
Year: 2010
Downloads: 96
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Abstract


Background and Objective: Gastric cancer is one of the world's most common malignancy, chemotherapy is the primary treatment of advanced gastric cancer. New anticancer drugs in recent years continue to emerge, chemotherapy combo update, but to improve the efficacy of advanced gastric cancer patients, the majority of patients is still not able to obtain effective control, one important reason for stomach cancer chemotherapeutic drugs resistance, and cause the failure of chemotherapy, however, currently used in clinical trials, most of the resistance reversal agents exist inefficient, high toxicity, change the pharmacokinetic issues such chemotherapeutic agents, to date, there is no ideal The drugs used in clinical overcome tumor resistance. Looking for efficiency, low toxicity natural drugs to overcome tumor resistance is one of the important topics of current cancer therapy research. Paeoniflorin the peony main active components, anti-inflammatory, immunomodulatory, hepatoprotective, neuroprotective function. In recent years, a small number of in vitro studies have reported confirmed paeoniflorin significantly inhibited the proliferation of liver cancer and lung cancer cells. Preliminary studies of our group showed that paeoniflorin inhibit gastric cancer cells (SGC-7901) proliferation and induce apoptosis in the resistant cell line has not been reported at home and abroad of the inhibitory effect of its role in the molecular mechanisms. Experiment resistant gastric cancer cells SGC7901/VCR research paeoniflorin on its mechanism of cellular resistance to explore the potential therapeutic effect of paeoniflorin resistant gastric cancer. Method: 1. Paeoniflorin alone or in combination with chemotherapy drugs that act on SGC7901/VCR cells after 48h, using methyl thiazolyl tetrazolium (MTT) method to detect cell proliferation inhibition rate; 2 was detected by flow cytometry (FCM) paeoniflorin alone or in combination with vincristine (VCR) apoptosis rate; 3 Western blot technology (Western Blot) analysis of different concentrations of paeoniflorin Bcl-2, Bcl-xl, Bax and nuclear NF-κB P65 protein expression of affect; 4. enzyme-linked immunosorbent assay (ELISA) to further define the transcriptional activity of NF-κB in the nucleus; 5 reverse transcription polymerase chain reaction (RT-PCR) detected before and after the application of paeoniflorin MDR1 mRNA expression changes; flow cytometry to detect different concentrations of paeoniflorin treatment of cells rhodamine 123 (Rh123) accumulation and discharge; 7. flow cytometry before and after the application of paeoniflorin cell surface protein P-glycoprotein (P-gp) expression . Results: 1. The paeoniflorin SGC7901/VCR cell growth significantly inhibited this effect in a dose-dependent manner; non-toxic concentration (80μg/mL) the paeoniflorin able to partially reverse the SGC7901/VCR cells to VCR and 5-FU resistance. 2 Paeoniflorin a concentration-dependent manner to promote apoptosis and to 80μg/mL the joint Paeoniflorin VCR than single drug VCR can significantly induce apoptosis. Western Blot and ELISA were confirmed paeoniflorin clear inhibition of NF-κB; With the increase in the concentration of paeoniflorin of NF-κB, Bcl-xl and Bcl-2 expression levels gradually reduced (P lt; 0.01), but Bax There is no significant impact. 4. Flow cytometry results presented paeoniflorin role of intracellular Rh123 fluorescence intensity was significantly enhanced; This results show that paeoniflorin after a significant increase in the concentration of intracellular chemotherapy drugs. 5. RT-PCR to detect the expression of mdr1 mRNA before and after the different concentrations Paeoniflorin applications no significant change (P gt; 0.01). Flow cytometry cell surface protein expression of P-gp results suggest the 40μg/mL Paeoniflorin P-gp protein expression did not affect (P gt; 0.01), but the 80 -320μg/mL paeoniflorin P-cut gp expression in a dose-dependent manner (P lt; 0.01). The Conclusion: paeoniflorin inhibit SGC7901/VCR cell proliferation and induce apoptosis; paeoniflorin part of multi-drug resistant reverse SGC7901/VCR cells reversed resistance mechanisms may be through the inhibition of NF-κB activation, thereby down the anti-apoptotic protein (Bcl-xl, Bcl-2) and P-gp.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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