Dissertation
Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology

The Study of Proliferation, Apoptosis and Its Mechanism Effect of Xanthorrhizol on Human Tongue Cancer Cell Line Tca8113 Cell

Author MaSai
Tutor DuanYuQin
School Hebei Medical University
Course Clinical Stomatology
Keywords Beam bone turmeric alcohols Tongue cancer Tca8113 cells Cell Cycle Apoptosis Bcl-2 Bax
CLC R285.5
Type Master's thesis
Year 2011
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Purpose: Tongue cancer yes oral and maxillofacial the Ministry of the most common malignant tumor of, in recent years in the world within the scope of its incidence rate year after year elevated, and its age of onset getting smaller and smaller, while its degree of malignancy is also increasingly high. Is easy to occurrence of Early cervical lymph node metastasis, and the metastasis rate a higher as the tongue cancer the main features of one of the. Currently tongue cancer the preferred treatment for way is to surgical treatment, the same time, preoperative, postoperative often need combined with chemotherapy in order to improve the efficacy, but the traditional chemotherapy is usually existence of forward to tumor cell drug resistance, chemotherapy drugs toxic side effects big in patients with is difficult to tolerate and so on affect the treatment the effect of unfavorable factors, so the research and development of treatment of malignant tumors new drugs, to become malignant tumor therapeutic area focus of the study. Currently, the domestic and foreign scholars more and more attach importance to From the flora and fauna, minerals, marine biological, and other natural substances medium Looking for efficiency and low toxicity of the treatment of malignant tumors of the drug, and has achieved certain achievements. Beam bone turmeric alcohol (Xanthorrhizol) yes from the turmeric genus Plants Indonesian Ezhu (C.xanthorrhiza) rhizome's volatile oil in the extracted out of the one kinds main ingredients, at present studies have shown, beam bone turmeric Alcohols on the cervical cancer, breast cancer and other malignant tumors has obvious inhibition. This issue through cultured in vitro tongue cancer Tca8113 cells, research bundles bone turmeric Alcohols on the tongue cancer Tca8113 cells proliferation and apoptosis of impact of, explore the beam bone turmeric alcohol in the presence tongue cancer the treatment's role mechanism for the tongue cancer of the chemical treatment of to provide experimental basis . Methods: The will recover after the's tongue cancer Tca8113 cells placed in 37 ℃, CO 2 saturated humidity 5% of the regular under the conditions of cultured, pending the cells to enter the logarithmic growth phase after the used for experiments. One flow cytometry (FCM) to detect cell apoptosis, cell cycle distribution: Collect different concentrations of beam bone turmeric alcohols role in 0h, 12h, 24h, 48h, 72h's tongue cancer Tca8113 cells and the control group cells in, cold PBS rinsing. Precooling 70% ethanol 4 ℃ fixed, upper machine detection ago centrifuged to remove fixative, with 0.5% pepsin digestion, propidium iodide (ProPidium Iodide, PI: 50mg / L Trinton X-100 1.0%) room temperature away from light staining. Flow cytometry on the machine testing, application software for analysis. 2 flow cytometry (FCM) detection of Bcl-2, Bax gene protein expression situations: collect treated with different concentrations beam bone turmeric alcohols role in 0h, 12h, 24h, 48h, 72h's tongue cancer Tca8113 cells and the control group cells in, according to conventional Methods for fluorescence labeled, flow cytometry fluorescence is detected intensity, in order to mean fluorescence intensity slopes at number indicates protein in expression amount of. Results: 1 bundle bone turmeric Alcohols on the tongue cancer Tca8113 cells cycle the impact of. 1.1 20μmol / L, 40μmol / L, 60μmol / L beam bone turmeric alcohols, respectively, role in Yu tongue cancer Tca8113 cells after 12h, each drug group S G2 / M phase cell cycle percentage of the with the control group of the compared to the, the difference was significant Xing (P lt; 0.01); drug action after 24h, each drug group S G2 / M phase cell cell cycle percentage of the compared with control group, the difference was significant Xing (P lt; 0.05); drug action after 48h, each drug group S G2 / M phase cell cell cycle percentage of the compared with control group, the difference was significant Xing (P lt; 0.05); drug action after 72h, each drug group S G2 / M phase cell cell cycle percentage of the compared with control group, the difference was significant Xing (P lt; 0.01). Each medication group and the only plus dimethylsulfoxide (DMSO) without the plus drugs's each solvent control group compared to the, the difference was significant Xing (P lt; 0.05). At different time points each use of drugs comparison between groups, in addition to medication after 72h, 20μmol / L group and the 40μmol / L between groups was no significant difference Xing outer (P> 0.05), the rest of at each time point in each group compare the differences were has the was significantly Xing ( P lt; 0.05). 1.2 each medication concentration groups role in different time S G2 / M phase cell cycle percentage of the's variance analysis showed that, 20μmol / L group medium, 48h with the 72h two point in time the comparison between, difference was no significant Xing (P> 0.05), the rest of at each time between the points, the difference was significant Xing (P lt; 0.05); 40μmol / L group medium, 48h with the 72h two point in time the comparison between, difference was no significant Xing (P> 0.05), the rest of each time point between, difference having a was significantly Xing (P lt; 0.05); 60μmol / L group, the each time point the comparison between, the differences were possess was significantly Xing (P lt; 0.05). 1.3 treated with different concentrations of drugs role in different time after a, G2 / M phase cell cycle percentage change in no obvious regularity. Two detected by flow cytometry beam of bone turmeric Alcohols on the tongue cancer Tca8113 cells cell apoptosis the impact of. 2.1 20μmol / L, 40μmol / L, 60μmol / L beam bone turmeric alcohols role in Yu tongue cancer Tca8113 cells after 12h, cell apoptosis rate compared with control group, the difference was significant Xing (P lt; 0.01); drug action after 24h , cell apoptosis rate compared with control group, the difference was significant Xing (P lt; 0.01); drug action after 48h, cell apoptosis rate compared with control group, the difference was significant Xing (P lt; 0.01); drug action after 72h, cell apoptosis rate compared with control group, the difference was significant Xing (P lt; 0.01); each medication group and the only plus dimethylsulfoxide (DMSO) without the plus drugs's each solvent control group compared to the, the difference was significant Xing (P lt; 0.01). Different time points each medication comparison between groups, in addition to medication after 12h, 20μmol / L group and the 40μmol / L between groups was no significant difference Xing outer (P> 0.05), the remaining time points in each group compare the differences were has the was significantly Xing (P lt; 0.01). 2.2 each medication concentration groups role in different time cell apoptosis in analysis of variance showed, in addition to 20μmol / L within the group 48h and the 72h comparison between, difference was no significant Xing outer (P> 0.05), the remaining each group each time point inter-comparison, differences were has the was significantly Xing (P lt; 0.05). 3 FCM detection of beam of bone turmeric alcohols role in Yu tongue cancer Tca8113 cells after the apoptosis-related gene protein Bcl-2, Bax the expression of. 3.1 Bcl-2 protein expression quantity: 3.1.1 12h, 24h group protein expression, respectively, Compared with control group, in addition to 20μmol / L outside the group, the rest differences in each group has the was significantly Xing (P lt; 0.05); 48h group protein expression and the control group compared to the, the difference was significant Xing (P lt; 0.05); 72h group protein expression compared with control group, the difference was significant Xing (P lt; 0.05); each medication group and the only plus dimethyl sulfoxide (DMSO ) rather than plus drugs's each solvent control group compared to the, the difference was significant Xing (P lt; 0.05); different time points each drug group comparison between, the difference was significant Xing (P lt; 0.05). 3.1.2 each medication concentration groups role in different time protein expression in analysis of variance showed, 20μmol / L within the group at each time point between the difference was not significant (P> 0.05), 40μmol / L group, in addition to 48h with the 72h between the was no significant difference Xing (P> 0.05) outside the, the rest at each time point difference among were significantly (P lt; 0.05), 60μmol / L group medium at each time point difference among were significantly (P lt; 0.05). 3.2 Bax protein expression quantity: Each medication concentration groups role in different time after the, in each group expression of Bax protein the amount of warp factorial design's variance analysis showed that, the time factor and dose factors that in-beam bone turmeric Alcohols on the cells expression of Bax protein the amount of the impact of middle, does not exist interaction effect (P> 0.05); one by one analysis of drug role in after the, the different times and different drug concentration pairs expression of Bax protein the amount of the impact of, results show that the experimental group and control group's difference was not possess was significantly Xing (P> 0.05). Conclusions: 1 bundle bone turmeric alcohols be able to significantly affect tongue cancer Tca8113 cells's cell cycle, so that cell cycle arrest in G0/G1 phase, so as to achieve anti-tumor purposes. The drug on the cell cycle blocking effect showed a time and dose dependent relationship. 2 beam bone turmeric alcohol in the presence certain range of concentration within a be able to showed a time, concentration-dependent manner induction of tongue cancer Tca8113 cells apoptosis in. 3 beam bone turmeric alcohols be able to significantly affect apoptosis-related gene protein of Bcl-2 expression, so that Bcl-2 protein expression gradually decreased, protein expression content in changing the showed a dose-dependent manner, the same time that the drug pairs of Bcl-2 protein expression in affect the in the reaches a certain concentration appear after time-dependent manner; of drugs on the apoptosis-related gene protein Bax the expression of no significant effect.

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