| Purpose:The correlation between TCM syndrome types and prognostic factor gene expression in breast cancer is currently the focus of research on the Combine traditional Chinese and western medicine in breast cancer.The classification of TCM syndromes in breast cancer is not yet uniform.In clinical practice,there are many types of syndromes based on the experience of physicians.The classification of different physicians is not the same.The physicians in the clinical treatment of different types of breast cancer differentiation,the lack of standardized standards,the understanding did not reach a consensus,the experience of summing up is not conducive to reference between each other.It is necessary to formulate new authoritative standards of TCM Syndrome type so as to improve the standardization of differentiation of breast cancer and facilitate the overall treatment of breast cancer by TCM.High expression of Cyclin D1 occurs in the early stage of breast cancer and persists in the process of infiltration and metastasis,and is closely related to the expression of ER.CCND1gene amplification is only about 20%in normal breast tissue,but over 50%of invasive breast cancer Cyclin D1 protein expression levels increased significantly,showing that Cyclin D1play an important role in the development of breast cancer.The expression level of Cyclin D1is significantly correlated with the degree of metastasis and proliferation of breast cancer.The overexpression of Cyclin D1 protein in ER-negative patients indicates early recurrence and poor prognosis.Further research on Cyclin D1 can provide a useful reference for the clinical treatment of breast cancer The purpose of this study was to investigate the differences in objective indicators between different TCM syndrome types,and to analyze the relationship between medical evidence and clinicopathological features and the expression of cyclin D1(CyclinD1),Provide objective evidence for clinical dialectical classification of breast cancer and guide clinical treatment.Material and method:Collected 100 patients with breast cancer who had undergone general surgery in the General Hospital of Shenyang Military Region from September 2016 to September 2017 and met the criteria for inclusion.Observed cases were all within 2 weeks before surgery.According to their clinical manifestations,the two deputy chief physicians dialectized into four types:liver qi stagnation,toxic heat accumulation,Qi blood deficiency group,and liver and kidney yin deficiency group.Liver-qi stagnation syndrome:single breast mass,or agglomerated as stone,with or without pain,two threatening pain,irritability,chest pain,bitterness,poor diet.Thin yellow or thin white fur with tongue,red tongue,and strong pulse string.toxic heat accumulation syndrome:Feeling upset or slightly hot,the breasts are red and hard,Frowning pain,stench,dry urine,dry mouth and throat,headache,insomnia,flushing,Red tongue with no moss,strong pulse slip.Qi blood deficiency syndrome:Dizziness and tinnitus,tiredness,fatigue,weight loss,shortness of breath,lack of complexion,Unresectable breast mass can cause breast agglutination,Color sluice,current sewage,The tongue is dull,the fur is thin,the pulse is thin and weak.Liver and Kidney Yin Deficiency syndrome:Menstrual disorders,accompanied by weak waist,dizziness,tinnitus,fatigue,Before the menstrual period,breast tenderness,breast swelling,or hard as stone,Crackling tongue,thin fur,narrow or weak pulse.The following pathological data were collected,histological types,primary tumor size,TNM stage,axillary lymph node metastasis,histological grade,and expression of immunohistochemical markers ER,PR,Her-2,and ki67,molecular subtype.Streptavidin-peroxidase-linked(SP)method was used to detect the qualitative expression of Cyclin D1 protein in cancer tissues and adjacent tissues.Immunoblotting(WB)was used to detect the quantitative expression of Cyclin D1 protein in cancer tissues and adjacent tissues.Real-time fluorescence quantitative PCR was used to detect the expression of Cyclin D1 in cancer tissues and adjacent tissues.SPSS 17.0 software was used for statistical analysis.Count data are expressed as percentages,χ2 test or Fisher’s exact probability method,and Mann-Whitney U test among the groups are used for analysis;measurement data are compared using one-way analysis of variance.The difference was statistically significant at P<0.05.Results:1.In the general data,the average age of patients with liver-qi stagnation group was49.86±12.559 years,and the average age of patients with liver-kidney yin deficiency group was 56.24±12.557 years.The average age of patients with liver-kidney yin deficiency group was greater than that of liver-qi stagnation group,with statistical significance(P<0.05).),The difference between the other groups was not statistically significant.There was no statistically significant difference in menstrual status between the groups at the time of initial diagnosis.2.In the pathological data,there was no significant difference in tumor size,lymph node metastasis,tumor stage,histological grade,ER,PR,Her-2,ki67 TCM syndrome groups.ER expression differs among the TCM syndrome types.In the liver-qi stagnation group,40(69.0%)cases were positive for ER,and 18(31.0%)cases were negative.ER-positive(12.5%)and negative(87.5%)cases were found in the toxic heat accumulation group.ER positive 8(61.5%)patients and negative 5(38.5%)patients in the Qi blood deficiency group.ER-positive 15(71.4%)patients and negative 6(28.6%)patients in the liver-kidney yin deficiency group.The chi-square test was performed on the expression of each group of patients.The results showed that:Χ~2=10.367,P=0.016<0.05,indicating that the expression differences of ER among the TCM syndrome groups were statistically significant.Each group was compared by Mann-Whitney U test,the rate of positive expression in toxic heat accumulation group was significantly lower than that in each group,and the other two groups were not statistically significant.The results of the relationship between each group of TCM syndrome types and molecular typing suggested that there were statistically significant differences between the four TCM syndrome types and the four types of molecular typing.Therefore,Luminal A is related to the syndrome of liver qi stagnation.3.The immunohistochemical results of Cyclin D1 expression in the TCM syndrome groups showed:There were 61 cases of Cyclin D1 positive expression in 100 cases of breast cancer specimens,42 cases of liver qi stagnation group were positive expression,the positive expression rate was 72.4%.The liver and kidney yin deficiency group showed positive expression in 9 cases,and the positive expression rate was 42.9%.The positive rate of liver qi stagnation group was significantly higher than that of liver and kidney yin deficiency group,and the result was statistically significant.The other two groups had no statistically significant difference.In terms of molecular subtypes,the positive rate of Luminal A was significantly higher than that of Her-2 overexpressed and basal cells,and the positive rate of Luminal B was significantly higher than that of Her-2 overexpressed and basal cells.4.Western blot results of Cyclin D1 expression in each group of TCM syndromes:The gray value of the blank control group was 346.78±274.29,the gray value of the liver qi stagnation group was 936.29±336.55,and the gray value of the toxic heat accumulative group was771.52±347.51.The gray value of Qi blood deficiency group was 798.35±319.02,and the gray value of liver and kidney deficiency group was 753.70±360.79.The protein expression level of CyclinD1 in each group was significantly higher than that in the blank control group(P<0.01).The protein expression level of CyclinD1 in the liver-qi stagnation group was significantly higher than that of the liver-kidney yin deficiency group in the TCM syndrome group(P<0.05).5.RT-PCR results of expression of Cyclin D1 in each group of TCM syndromes:The expression of CyclinD1 mRNA was significantly increased in the liver-qi stagnation group and the Qi blood deficiency group compared with the adjacent tissue.There was no significant difference in the expression of CyclinD1 mRNA between the liver-kidney yin deficiency group,the toxic-heat accumulation group and the adjacent tissue.The expression of CyclinD1 mRNA was significantly increased in the liver qi stagnation group compared with the liver-kidney yin deficiency group and the toxic heat accumulation group.There was no significant difference in the mRNA expression level of CyclinD1 between the other groups.Conclusion:1.The dialectical diagnosis of breast cancer is divided into liver qi stagnation,toxic heat accumulation,Qi blood deficiency,and liver-kidney yin deficiency.Four types are in line with clinical practice.Age is the influencing factor of TCM syndrome type classification,and young people often have liver-qi stagnation syndrome types.Elderly and postmenopausal patients often have liver kidney yin deficiency syndromes;2.ER positive patients were mostly with liver qi stagnation syndrome,blood deficiency syndrome,liver-kidney yin deficiency syndrome,and ER negative was mostly toxic heat accumulation syndrome type;3.The percentage of overexpression of Cyclin D1 protein in liver-qi stagnation syndrome was significantly increased,the proportion of Luminal A was the highest in the liver-qi stagnation syndrome,CyclinD1 was over-expressed in the molecular class Luminal A,and the liver-qi stagnancy syndrome was associated with over-expression of CyclinD1.Overexpression of CyclinD1 may be the molecular mechanism of liver qi stagnation;4.The expression of CyclinD1 protein in liver qi stagnation group was higher than that in liver and kidney yin deficiency group.The expression of CyclinD1 mRNA was up-regulated in liver qi stagnation group and blood deficiency group,which was not completely consistent with the protein expression,suggesting that there may be other factors involved in the regulation of CyclinD1 protein expression. |
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