Investigation Of Expression Of Tumour Necrosis Factor And Its Receptor In Yin-deficiency Syndrome Of Cancer

Cancer has become one of the leading causes of death. The incidence and death rate is still increasing in a number of countries. The treatment effect of cancer is low, because 60%～70% of them are at advanced stage when final diagnosed. So it is important to study alleviative treatment. However, many patients with cancer of advanced stage may accompany Yin-deficiency syndrome, which will decrease their immunity and influence the life quality of the patients. If we can find the correlation between cancer and Yin-deficiency syndrome, investigate the essence and pathogenesis of Yin-deficiency syndrome, understand the molecular pathophysiological mechanism of the occurrence of Yin-deficiency syndrome, we will raise the treatment effect of Traditional Chinese Medicine(TCM) and improve the life quality of patients.Prof. WeiXi Shen and Yan Sun based on the theory and method of molecular biology, propose that the essence of Yin-deficience syndrome can be cytokines, as its pathogensis can be due to the perturbation of cytokines networks characterized by the elevation of the gene expression and the biology activity of Interleukin-1(IL-1), Tumour Necrosis Factor(TNF) and so on which results in the turbulence of cytokine networks.ObjectiveWe have used the technology of gene chip and immunohistochemistry to investigate the changing regulations of TNF and its receptors, to verify the preliminary conclusion that the pathogenesis of the Yin-deficiency syndrome is due to the disequilibrium status of cytokine networks resulted from the relative increase of the bioactivity of the pro-inflammatory cytokines like IL-1, TNF and so on.Methods1. To determine the gene expression of TNF superfamily and its recepters of Yin-deficiency syndrome of cancer by Affymetrix Gene Chip.To select 3 cases of cancer patients with Yin-deficiency syndrome and 3 cases of non-Yin-deficiency syndrome. Patients of Yin-deficiency syndrome: one case of lung cancer, one case of colon cancer and one case of gastric cancer. Patients of non-Yin-deficiency: one case of lung cancer, one case of colon cancer and one case of gastric cancer. 2ml blood samples were collected from the patients. The total RNA which was abstracted from the blood was reversely transcribed to cDNA to prepare the hybridization probes. After washing the gene chip, the cDNA microarray was scanned for the fluorescent signals. The data were analyzed by Gene Chip Operating Software. The signals were ordering by the method of Normalization and the diferences between the two patients were showed.2. To determine the expression of tumor necrosis factor alpha(TNF-α) and tumor necrosis factor receptor I (TNFR I) of Yin-deficiency syndrome of lung cancer by immunohistochemistry.To select 20 cases of lung cancer patients with Yin-deficiency syndrome and 20 cases of lung cancer patients with non-Yin-deficiency syndrome. They were divided into two groups: Yin-deficiency group and non-Yin-deficiency control group. All the samples were resected from patients who were operated. In this paper, the expression of TNF-αand TNFR I were studied by immunohistochemical method. The data were analyzed by SPSS11.0 statistical software, x~2 test was carried out to determine if the change was statistically significant.α=0.05 was considered statistically significant.Results1. The Gene Chip results showed: The expression level of TNF-α, TNFR I , tumor necrosis factor receptor II (TNFRII), tumor necrosis factor receptor superfamily, member 10(TNFRSF10), tumor necrosis factor receptor superfamily, member 13b(TNFSF13b) in lung cancer Yin-deficiency patient is higher than in lung cancer non-Yin-deficiency patient. But the expression level of tumor necrosis factor alpha-induced protein 8(TNFAIP8) in lung cancer Yin-deficiency patient is lower than in lung cancer non-Yin-deficiency patient.The expression level of TNF-α, tumor necrosis factor beta(TNF-β), tumor necrosis factor superfamily, member 8(TNFSF8), tumor necrosis factor superfamily, member 10(TNFSF10), TNFSFBb, tumor necrosis factor alpha-induced protein 6(TNFAIP6), tumor necrosis factor receptor superfamily, member 17(TNFRSF17), tumor necrosis factor receptor superfamily, member 25(TNFRSF25) in colon cancer Yin-deficiency patient is higher than in colon cancer non-Yin-deficiency patient. But the expression level of TNFR I , TNFR II, tumor necrosis factor superfamily, member 5(TNFSF5), TNFAIP8 in colon cancer Yin-deficiency patient is lower than in colon cancer non-Yin-deficiency patient.The expression level of TNF-α, TNF-β, TNFR I , TNFR II in gastric cancer Yin-deficiency patient is higher than in gastric cancer non-Yin-deficiency patient. But the expression level of TNFSF5, TNFRSF10, TNFSF13b, TNFAIP6, TNFAIP8, TNFRSF17 in gastric cancer Yin-deficiency patient is lower than in gastric cancer non-Yin-deficiency patient.2. The immunohistochemical results showed: In the lung cancer cells and other normal lung cells, the expression level of TNF-αin Yin-deficiency group were obviously higher than that in non-Yin-deficiency control group(P<0.05). The difference were significant. The difference of expression level of TNFR I between Yin-deficiency group and non-Yin-deficiency control group were not significant (P>0.05).Conclusions1. Our research preliminarily verifies the correlation between TNF and Yin-deficiency syndrome and verifies the theoretical conclusion that the essence of Yin-deficiency syndrome is cytokines. The pathogensis can be due to the gene expression level and the 4bioactivity of pro-inflammatory cytokines such as TNF-αincreased absolutely or relatively, though the soluble cytokine receptors couldn't antagonise effectively. As a result, the balance of cytokine networks was destroyed and the patient appeared a series of symptoms and signs of Yin-deficiency syndrome.2. The essence and pathogensis of Yin-deficiency syndrome has a certain heterogeneity: the pathogensis of Yin-deficiency syndrome has a bit difference in different patients of different cancer, so the gene expression of TNF and its receptors also has differences in different patients.3. We suppose that the pathogensis of Yin-deficiency syndrome is that: through certain signal transduction pathway, the transcription factor like NF-κB and so on is activated which provokes the gene expression of cytokines like TNF and so on. The elevation of the contents and the biology activity of TNF and other cytokines cause the perturbation of cytokine networks and cause a series of symptoms and signs which we call Yin-deficiency syndrome.