Dissertation > Excellent graduate degree dissertation topics show

Research on the Molecular Mechanism of Blood-stasis-removing and Phlegm-resolving Therapy’s Resistance to the Lung Cancer Metastasis

Author: JiangLin
Tutor: KongQingZhi
School: Hubei University of Chinese Medicine
Course: Chinese medical science
Keywords: the blood-stasis-removing and phlegm-resolving therapy lungcaner tumor metastasis blood stasis syndrome phlegm syndrome
CLC: R734.2
Type: PhD thesis
Year: 2012
Downloads: 278
Quote: 0
Read: Download Dissertation

Abstract


Research Background and Objective:Lung cancer is one of the malignant tumors with the highest rate of incidence and mortality, and80-90%death of patients with lung cancer is caused by the metastasis. Invasion&metastasis is one of the essential characteristics of malignant tumors, but also the major cause for the treatment failure and mortality.Therefore, the metastasis control is a key factor to determine the prognosis of cancer patients. In recent years, the studies of tumor invasion and metastasis have made great progress both at home and abroad, they have clarified some of the key points and mechanism of tumor invasion and metastasis, and have developed several kinds of drugs to prevent diversion, but so far no drugs have been widely applied in the clinical prevention of tumor metastasis.Chinese medicine has a long history and an important position in the treatment of tumors. In the long-term medical practice, traditional Chinese medicine formed a unique theoretical system of prevention and treatment of malignant tumors. The blood-stasis-removing and phlegm-resolving therapy is an important method for the treatment of malignant tumors, especially for the lung cancer treatment.Modern experimental studies have shown that controversies still exist over the anti-tumor and metastasis effects of the medicines for blood-stasis removing and phlegm resolving. Thus the exploration of the molecular mechanism of blood-stasis-removing and phlegm-resolving therapy’s resistance to the lung cancer metastasis is practically significant.This study was conducted under the guidance of traditional Chinese medicine theory, and followed the principle of the combination of the syndrome differentiation and disease differentiation, and combination of macro syndrome and microcosmic syndrome.We conducted in vivo and in vitro experiments with the blood-stasis-removing therapy (Taohong Siwu decoction) and phlegm-resolving therapy (Erchen decotion) to study the classical mouse Lewis lung carcinoma metastasis model and human highly metastatic lung carcinoma95-D cells, and to explore the molecular mechanism of blood-stasis-removing and phlegm-resolving therapy’s resistance to the lung cancer metastasis.Meanwhile, we applied the idea of disproval of the’syndrome and therapy’ in Traditional Chinese Medicine (TCM) to the attempt at searching for the biological essence of lung cancer with blood stasis syndrome and phlegm syndrome, offering positive method and reference to the innovation and modernization of TCMMaterials and Methods:The first section:Experiments in vivo1、Building mouse Lewis lung carcinoma metastasis model2、Animal models were randomly divided into eight groups,10mice per group:saline control group, high-dose blood-stasis-removing and phlegm-resolving therapy group, low-dose blood-stasis-removing and phlegm-resolving therapy group, high-dose blood-stasis-removing therapy group, low-dose blood-stasis-removing therapy group, high-dose phlegm-resolving therapy group, low-dose phlegm-resolving therapy group, and cisplatin group.3Medications were respectively given to the8groups for21days. Periodic measurements of changes in the transplanted tumor’s volume and mice’s weights in each group were conducted during the experiment. After21days, mice in each group were killed and weighed, then, tumor masses were taken off and weighed for calculating the inhibition rates of the experimental groups and histopathological section of transplanted tumor were made.4The lungs of mice were taken to calculate the number of metastases tumor nodules on lung surface, and then, the inhibition rates of lung metastasis in experimental groups were calculated. The histopathological sections of lungs were made to observe the pathological changes in lungs.5With the SABC immunohistochemical method, detection of expression of CD31in Lewis lung carcinoma transplanted tumor was conducted and the micro vessel density (MVD) of the tumor in tumor-bearing mice is calculated.6The expressions of molecules (TF and MMP-2and VEGF in E-cad, CD44) related to transplanted tumor in Lewis lung carcinoma on protein level was detected with SABC immunohistochemical method.The second section:Experiment in vitro^Preparations for the serums containing the blood-stasis-removing and phlegm-resolving therapy or blood-stasis-removing therapy or phlegm-resolving therapy were made. Human highly metastatic95-D cell were cultivated in vitro.2,The impact that the blood-stasis-removing and phlegm-resolving therapy might have on human lung cancer95-D cell proliferation was detected with MTT method.3, Adhesive analysis experiment was conducted to detect the effect that the blood-stasis-removing and phlegm-resolving therapy might have on the human lung cancer95-D cell’s homotypic adhesion and on the heterotypic adhesion to human umbilical vein endothelial cells (HUVEC).4, Wound healing experiment was applied to the detection of the impact on the human lung cancer95-D cell’s migration made by the blood-stasis-removing and phlegm-resolving therapy5, Transwell invasiveness method was used to detect the impact that the blood-stasis-removing and phlegm-resolving therapy might have on human lung cancer95-D cell.6, The impact on the expression of mRNA of the molecules (TF and MMP-2and VEGF in E-cad, CD44) related to transplanted tumor in human lung cancer95-D cells made by the blood-stasis-removing and phlegm-resolving therapy was investigated by the real-time PCRResultsThe first section:Experiment in vitro1, Weight changes of the mice in every group:except the cisplatin group, the weights of the mice in each group increased gradually. The overall trend of weight in the cisplatin group was that it deceased first and that it tended to increase later, but the weight hasn’t returned to that of the beginning of the experiment till the end.2, Changes in the volumes of transplanted tumors in each group:volume of transplanted tumor in saline control group was the fastest-growing one, followed by high-dose blood-stasis-removing and phlegm-resolving therapy group and cisplatin group.3, The tumor inhibition rate in each group:the rates of the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups were38.02%and24.11%respectively and those of the high-and low-dose blood-stasis-removing therapy groups were19.28%and14.25%, while the high-and low-dose phlegm-resolving therapy groups got the inhibition rates of10.74%and4.96%, presenting certain dose-dependent manner. The inhibition rate in Cisplatin group was59.22%.4, Lung metastasis inhibition rates in each experimental group:the Lung metastasis inhibition rates in the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups were39.77%and29.25%and those of the high-and low-dose phlegm-resolving therapy groups were20.92%and16.42%, and it also presented certain dose-dependent manner. The lung metastasis inhibition rates in high-and low-dose the blood-stasis-removing therapy groups were11.91%and20.69%respectively.5, The effect on micro vessel of the transplanted tumor in the every experimental group:high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups, high-dose blood-stasis-removing therapy and high-dose phlegm-resolving therapy groups could inhibit the angiogenesis. Whereas the inhibitory effects of the low-dose blood-stasis-removing therapy group and phlegm-resolving therapy group were not conspicuous.6, The expressions of molecules (TF and MMP-2and VEGF in E-cad, CD44) related to transplanted tumor on protein level:both the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups could improve the expression of E-cadherin of transplanted tumor in lung cancer and lower that of the CD44, MMP-2, VEGF and TF.7, The high-and low-dose blood-stasis-removing therapy groups mainly reduced the expression of TF, MMP-2and VEGF in transplanted tumor in Lewis lung carcinoma; The high-and low-dose phlegm-resolving therapy groups mainly enhanced the expression of E-cadherin whereas lessened that of CD44.The second section:Experiment in vitro 1, Inhibition rates for95-D cell proliferation in each experimental group: In the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups, the rates were31.49%(24h),36.35%(48h) and25.32%(24h),28.14%(48h), and those of high-and low-dose blood-stasis-removing therapy groups were15.98%(24h),21.96%(48h) and10.60%(24h),19.72%(48h), and in the high-and low-dose phlegm-resolving therapy groups, the rates decreased to4.43%(24h),10.34%(48h) and0.79%(24h),7.36%(48h)2, The impact on the95-D cell’s homotypic adhesion in each experimental group:the95-D cell’s homotypic adhesion was improved in the low-dose blood-stasis-removing and phlegm-resolving therapy group and the high-dose phlegm-resolving therapy groups (P<0.01), while the high-dose blood-stasis-removing therapy group reduced the95-D cell’s homotypic adhesion (P<0.05).3, The impact on the95-D cell’s heterotypic adhesion in each experimental group:The high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-and low-dose phlegm-resolving therapy groups restrained the adhesion of95-D and HUVEC (P<0.01), and there existed the certain dose-effect relationship. Compared with the blood-stasis-removing therapy group and the phlegm-resolving therapy group, the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups had a stronger effect on inhibiting the adhesion.4, The impact on the migration of95-D cell in each experimental group: the high-dose and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-dose blood-stasis-removing therapy and phlegm-resolving therapy groups inhibited the migration of95-D cell (P<0.05, P<0.01). 5, The impact on the invasiveness of the95-D cell in each experimental group:the high-dose and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-dose blood-stasis-removing therapy and phlegm-resolving therapy groups all repressed the invasiveness of95-D cell (P<0.05, P<0.01).6, The Impact on the transcription of genes(TF、MMP-2、VEGF、E-cad. CD44) related to the95-D cell tumor metastasis:1), The expression of TF mRNA in95-D cell was inhibited in the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-dose blood-stasis-removing therapy and phlegm-resolving therapy groups(P<0.01). The low-dose phlegm-resolving therapy group facilitated the expression of TF mRNA in95-D cell (P<0.01).2), The high-dose blood-stasis-removing and phlegm-resolving therapy group and the low-dose blood-stasis-removing therapy group inhibited the expression of MMP-2mRNA in95-D cell (P<0.01). The high-dose blood-stasis removing therapy promoted the expression of MMP-2mRNA in95-D cell (P<0.01)3), The expression of VEGF mRNA in95-D cell was inhibited in the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-and low-dose blood-stasis-removing therapy groups and the high-and low-dose phlegm-resolving therapy groups (P<0.01), and there existed certain dose-dependent manner.4), The expression of E-cad mRNA in95-D was improved in the high-and low-dose blood-stasis-removing and phlegm-resolving therapy groups and the high-and low-dose blood-stasis-removing therapy groups and the high-and low-dose phlegm-resolving therapy groups (P<0.01). The low-dose therapy groups, however, presented more obvious effects than high-dose groups. 5),The expression of CD44mRNA was improved in low-dose blood-stasis-removing and phlegm-resolving therapy and low-dose blood-stasis-removing therapy and low-dose phlegm-resolving therapy groups (P<0.01). The expression of CD44mRNA was inhibited in the high-dose blood-stasis-removing and phlegm-resolving therapy group (P<0.01).Conclusion1, The blood-stasis-removing and phlegm-resolving therapy can inhibit the growth and metastasis of lung cancer, and have different degrades of effects on the tumor cell proliferation which is the key point in metastasis of lung cancer, on adhesion of tumor cell, on the degradation of extracellular matrix, on the migration and invasion of tumor cell, and on tumor angiogenesis.2, Blood-stasis-removing therapy and phlegm-resolving therapy not act on every step during the tumor metastasis, and different therapy may act on different steps of the metastasis. Besides, the same therapy exerts dissimilar function in different steps of the metastasis:it inhibits the metastasis in some steps while improves in others. On the whole, however, the blood-stasis-removing and phlegm-resolving therapy inhibits the growth and metastasis of lung cancer.3,The mechanism of the blood-stasis-removing and phlegm-resolving therapy’ resistance to the lung cancer metastasis probably is that the therapy affects the expressions of TF, MMP-2, VEGF, E-cad and CD44in lung tumor cells on the level of protein synthesis and gene transcription, thus impacting on the proliferation, adhesion, migration and invasion of tumor cell, and on the tumor angiogenesis and degradation of extracellular matrix. The therapy can inhibit the lung tumor metastasis through multi-process, multi-step, multi-target. 4,The essence of lung cancer with blood-stasis syndrome may lie in the molecular network formed by abnormal expressions of TF, VEGF and MMP-2in lung tumor cells.5,The essence of lung cancer with phlegm syndrome may be the molecular network formed by the abnormal expressions of adhesion molecules such as E-cadherin and CD44.

Related Dissertations

  1. Preliminary Studies on Peptidomics and Metabonomics of Cerebral Infarction with Blood Stasis Syndrome,R277.7
  2. The Regulate and Controlof Slug on P53-MDM2 System,R730.2
  3. α1, 3 fucosyl transferase Ⅳ (Fut4) upregulated increased metastatic potential of squamous cell scc12,R730.2
  4. CXCR1/CXR2 receptor antagonists on mouse ascites hepatoma lymphatic metastatic potential impact,R735.7
  5. Construction of Mouse CXCR3A and Human CXCR3B Gene Transfected Cell Lines, Identification and Study of Their Biological Functions,R730.3
  6. Research of Gene Expression in Side Population (SP) Cells of Neuroblastoma,R739.4
  7. Experimental Study of the Treatment to the Syndrome of the Blood Stasis Due to Cold Accumulation of Primary Dysmenorrheal by Ningxin Huantong Tang,R285.5
  8. Clinical Study on Treatment of Nonliquefaction of Semen Which is Damp Heat with Blood Stasis Syndrome by Shengjingcuhuatang,R256.56
  9. The Intervention Research of hU-Ⅱ、TM on Coronary Artery Spasm Patients with Wen YangHuoXue Method,R259
  10. The Clinical Research of Pinggan Xifeng Huayu Tongluo Treatment Migraine (Liver Wind Carry Blood Stasis Syndrome ),R277.7
  11. Sankuhuoluoyin Chinese Traditional Medicine to the Clinical Curative Effect of the Treatment of Coronary Heart Disease, Angina (Heart Blood Stasis),R259
  12. Construction and Expression of Fusion Gene (RGD)3 - DAB391 Prokaryotic Expression Vector and Study of Its Tumor Depression Effect in Vitro,R737.9
  13. The Importance of Factor Phlegmatic Hygrosis in the Type Ⅱ Diabetes Combine with Cerebral Infarction, and Its Associativity with Leptin, Adiponectin,R743.3
  14. The Study of the Characters of the Symproms of the Blood Stasis Syndrome and the Qualty of Life in Hypertensive Patients with Abnormal Blood Fat,R259
  15. Relevance Study between Blood Stasis Syndrome of Mid and Late NSCLC Patient and Relevant Symptoms and Portion of Laboratory Index,R273
  16. Relativity Research on the Correlation of the Serum Bilirubin and Lipid Levels with the Phlegm and Blood Stasis of TCM Syndrome in Patients Coronary Heart Disease,R259
  17. The Clinical Study is about the Research of Efficacy for Treating Diabetic Peripheral Neuropathy (DPN) with Tonifying Qi Huoxue Away Bi-complex Soup,R285
  18. Eye Signs of Blood Stasis Syndrome and Vascular Endothelial Cell Injury in the Experimental Study,R255.7
  19. Non-ST- segment elevation acute myocardial infarction GRACE score and blood stasis syndrome,R259
  20. Relationship Between Carotid Atherosclerosis and Phlegm-blood Stasis Syndrome of Essential Hypertension,R277.7
  21. Expression and Significance of KiSS-1、NF-κBp65 and MMP-9 in Non-small Cell Lung Cancer,R734.2

CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
© 2012 www.DissertationTopic.Net  Mobile