The Basic And Clinical Translation Research Of Anti Breast Cancer Hematogenous Metastasis With The Traditional Chinese Medicine Treatment Under The Law "Yiqi Huoxue"

Breast cancer is a malignant tumor that seriously threat women’s lives and health. The metastasis of breast cancer is one of the direct causes which deprivate patients’lives. Traditional Chinese Medicine has curative effect in anti breast cancer metastasis. We find that there is a closely relationship between Xueyu syndrome and hematogenus in breast cancer disease basing on the reviewing of literature reports. Also there is large academic disputation in using of Huoxue huayu herbs in cancer disease:how to use, the dosage and the mechanism haven’t been explicated. In the past half century, my mentor Professor Sun Guizhi has beening treated malignant tumor patients with integrative oncology medicine methods and has accumulated rich experience of breast cancer treatment. She points out that Qixu, Xueyu, Duning are the fundamental pathogenesis of breast cancer invasion and metastasis, she found that the performance of Xueyu in breast cancer not only has the common characters in all the tumor disease, but also has it’s own specialty which based on the meridian patrolling and the situation of Qixue in the meridian. According to the results of basic research funded by national science foundation combined with the experience, Professor Sun Guizhi designed a formula for Xueyu symptom of breat cancer patients which contains five herb medicines:Huangqi, Sumu, Fengfang, Puhuang, Paoshanjia. Under the guidance of my mentor’s academic theory in anti breast cancer metastasis, this deviation, we force on efficacy, mechanism and usage of Traditional Chinese Medicine during the deep and systematic research.1PurposeThe research of my mentor’s academic theorySummarize professor’s experience in breast cancer treatment.Basic researchGain Yiqi huoxue core formula’s best compatibility of "Yiqi huoxue formula" and it’s appropriate clinical dose. Investigate the scientific connotation of mentor’s academic theory in the two major area at the point of anti breast cancer hematogenesis:Epithelial to mesenchymal transition and Angiogenesis.Clinical translational researchFormulate the quantify diagnostic criteria of Xueyu Syndrome as the converhence of theoretical and basic research which provide technical support for mentor’s clinical academic theory.2MethodThe research of mentor’s academic theorysummarize professor Sun Guizhi’s experiences basing on the experience of learning clinical skill under Professor’s guidance in the outpatient.Basic researchUsing the tumor bearing mouse model, filter the "Yiqi huoxue core formula" which has the strongest activation in anti breast cancer proliferation in vivo; Using drug containing serum pharmacological method, gain the appropriate dose of core formula. Using the methods of vitro and in vivo, explore the molecular mechanism of core formula’s function in anti breast cancer hematogenes metastasis via the regulation of TGF-β/Smad pathway mediated epithelial to mesenchymal transition and angiogenesis.Clinic translational researchUsing clinical epidemiology and statistical methods, study the quantify diagnostic criteria of Xueyu syndrome in breast cancer and automate the process of discrimination by JAVA.Test the differences of TGF-β level in breast cancer patients’ serum with "Xueyu syndrome" or not. Converge with the basic research.3.Result3.1The experience of Professor Sun Guizhi in treating breast cancersThe main features of the mentor:treating disease according syndrome diagnosis, choosing famous formula according with the exact syndrome, different kinds of disease different treatment, combining the disease with syndrome, carefully choosing herbs according to the results of base medical researchs, emphasizing the meridian and returning to the tradition. And Fuzhegn, Shugan, Liqi, Huoxue, Jiedu are the key points in professor’s treatment experience. Her treatment of the lung, bone, brain, liver metastasis of breast cancer often refers to the physiological characteristics of the organ itself. Obeying these distinctive laws, the treatment efficacy usually becomes outstanding.3.2Basic research3.2.1The best combination study of Yiqi huoxue formula by apart prescription methodWe find that the core Yiqi huoxue formula consists of "Huangqi, Sumu, Fengfang, Paoshanjia" has the strongest activity in preventing EMT6cells proliferation in vivo, the difference with control group has statistical significance, P<0.05.3.2.2The appropriate dose of Yiqi huoxue formula reseachThe formula dose "Huangqi30g, Sumu6g, Fengfang6g, Paoshanjia6g" and the2times dose can inhibit the MCF-7well proliferation, especially the2time dose serum containing herb drugs has more powerful activity.4times dose has the inhibiton tendency, but the differences has no statistical significance. At the72hours point, the6dose serum has the activity of promote the cell proliferation, P<0.05. At the24,48,72hours point,9dose serum will promote the cell proliferation, P<0.05.3.2.3The anti mouse breast cancer cell4T1metastasis to lung research under the treatment of Yiqi huoxue core formulaThe model used in this research in BALB/C mouse bearing4T1breast cancer cells. We found that the Yiqi huoxue core formula can prevent4T1cells’metastasis to lung. The inhibitory rate was58.82%, less than the Taxel group whose inhibitory rate was65.88%. But the differences between formula group and Taxel group has no statistical significance.3.2.4The basic research of Yiqi huoxue formula’s function in anti breast cancer hematogenous metastasis3.2.4.1The basic research of Yqi huoxue formula’s function in preventing epithelial to mesenchymal transition progress of beast cancerThe Yiqi huoxue formula could prevent the EMT process in MCF-7cells which were under going the EMT induced by2ng/ml TGF-β. We checked the preventive function at the protein level and the transcription level by identified the changes of several EMT markers and the difference ability of invasion and migration of different treated cells. These results well supported the Yiqi huoxue formula’s anti metastasis function at the prevention the deterioration process of breast cancer cell itself.3.2.4.2The research of Yiqi huoxue formula function in inhibiting the TGF-β activit during the TGF-β induced EMT processWe use dual reciferase report assay combined with real time PCR technology to identify theSmad molecular’s entering cell nucleus process, down regulated the activity of TGF-β/smad pathway, reduced the expression of transcription factor SNAIL.The TGF-β level in the4T1breast cancer cells bearing mouse was low than that in the control group. And the differences has stastistical significance. But this type of function is some lower than the Taxel. The conclusion was that Yiqi huoxue formula may inhibit the EMT progress via down regulating the TGF-β level in serum.3.2.4.3the research of anti breast cancer angiogenesis function of Yiqi huoxue core formulaWe compared the angiogenesis situation in the edge of the lesion and the internal location of the tumor tissue coming from the4T1breast cancer cell bearing mouse via the frozen biopsy organization immunofluorescence techiques. We found that the Yiqi huoxue fonnula had the inhibitory effects of angiogenesis in both the edge and the internal location. Real time PCR examination claimed the this function may be associated with the down regulation of VEGF-A.3.3Clinic transflational researchQuantify diagnosis criteria is established. The diagnostic threshold is16point, whose sensitivity is95.0%, specificity is81.8%, accuracy is90.3%, positive likelihood ratio is5.225, negative likelihood ratio is6.11%. The test performance is excellent.The TGF-β level in breast cancer patient with "Xueyu syndrom" is higher than that with out "Xueyu syndrom",P<0.05. 4ConclusionsThe "Yiqi huoxue core formula" based on Professor Sun Guizhi’s clinical experience has the function of anti-breast cancer hematogenes metastasis via the inhibition of molecular Smads’entering nuclear and binging to the promoter area of target genes which can prevent the EMT process of breast cancer. Also the prevention of angiogenesis is it’s mechanism."Yiqi huoxue core formula" can be used in breast cancer cases at the recommended dose referring to the breast cancer’Xueyu syndrome’Professor SunGuizhi’s experience of treating breast cancer hematogenes metastasis under the law of ’Yiqi huoxue’ owns rich theoretical content and scientific connotation which is worthing learning, inheritance, research and development.4The main innovation4.1Reveal the scientific connotation of "Yiqi huoxue" law in preventing breast cancer hematogenes by the research of anti breast cancer cell proliferation and angiogenesis in vitro via the bioinformation and molecular biological techniques. Provide new ideas in inheriting and mining clinical experience of famous Chinese medicine master.4.2The increased TGF-β level is similar with that in the serum of breast cancer patients."Yiqi huoxue core formula" can decrease the TGF-β level in the serum of tumor bearing mouse model which afford new basis for real time observation and drug intervention treatment in clinical breast cancer cases.The established quantify diagnostic criteria of breast cancer " Xueyu syndrome" with the measuring statistical analysis techniques afford clinical conversion basis and practical tools for the promotion of famous Chinese medicine masters’ experience.