Promoting Blood Circulation And Proliferation Of Drugs On PG Cells Under Hypoxia Effects Of Invasion And Metastasis

Metastasis is the substantive characteristic in the tumor biological behavior and it is also one of the major causes of death in cancer patients, so, it is important to study the metastasis so as to control and cure tumor. The theories of traditional Chinese medicine conclude the pathogenesis of malignancy into three points:qi stagnation and blood stasis, retention of water-dampness and accumulation of phlegm-dampness, qi and blood asthenia. Many experiments and clinical researches show that almost every tumor patient is in HYPERCOAGULABLE STATE. The blood of tumor patient who is in HYPERCOAGULABLE STATE is thicker and sticker than normal. And it is known as Tumor Blood Stasis Syndrome. The HYPERCOAGULABLE STATE exists in four pathological staging:invasion, proliferation, infiltrates and metastasis. The HYPERCOAGULABLE STATE will be more obvious as the tumor is in the bad way. On the basis of therapeutic principles, blood-activating drugs are used widely to the therapy of tumor. However some recent clinical and basic experimental researches found that some kind of blood-activating drugs may promote the tumor metastasis. And there are also some parts of the researches found that blood-activating drugs can inhibit tumor proliferation and metastasis. So what is the blood-activating drugs' effect in cancer treatment process? And what is the mechanism of its function? It is difficult to make a conclusion accurately due to there are various blood-activating drugs and different agents. According to my teacher's research, we find that blood-activating drugs can affect the tumor cell's metastasis based on different conditions to show inhibition or promotion. Due to its complexity, we have conduced further researches.This research project is set out on the basis of our tutor's precedent researches and is supported by National Natural Science Fund. On the basis of our early stage research work, we used Sappanwood and Ligusticum wallichii Franch, prepared Rat serum contained drugs, pretreated PG cells, then found that the serum containing can promote PG cell proliferation. All things considered, we have chosen TAN II A as intervention drugs. The in vitro experiment is divided into normoxia state and hypoxic state. And methods of Flow Cytometry and immunohistochemistry were used to measure the relative index of CD44s,CD44v6,HIF-1α,CD29,CD324 and so on. We have reconstructed experiment about invasion of the basement membrane to show TANⅡA's affect on PG cell's proliferation and adhesion potential. Effects of the serum containing which were processed with Sappanwood and Ligusticum wallichii Franch on PG cell's proliferation:there are differences (p<0.05) between the results in normoxia state and hypoxic state. And this is true of the serum containing(p=0.016<0.05). The serum containing can promote PG cell's proliferation (20% serum should affect PG cell's proliferation most).Effects of TANⅡA on PG cell's proliferation:there are differences between the results (p<0.05) in normoxia state and hypoxic state. And this is true of the concentration of TAN II A (p<0.05). TANⅡA is good for PG cell's proliferation. The PG cell's proliferation increased with the time prolonging of TAN II A's effect(it will affect the proliferation most when lasting to 72 hours). The hypoxic state is good for PG cell's proliferation.Determine the expressions of CD29 and CD324 by the FCM:CD29:TANⅡA can reduce the expressions of CD29 and inhibit cell's metastasis as the concentration of TANⅡA increasing in normoxia state and hypoxic state.CD324:it will increase the expressions of CD324 and promote PG cell's proliferation in modest hypoxic state, so it is good for the invasion of the PG cells. The expressions of CD324 will increase as the concentration of TANⅡA increasing in normoxia state while it will decrease in hypoxic state.The expressions of HIF-lα, CD44s and CD44v6 were assessed by immunohistochemistry:The expression of CD44s will be higher in normoxia state than in hypoxic state, while The expression of CD44s will the same in normoxia state and hypoxic state. This result shows that PG cell's metastasis can be inhibited in normoxia state. in the basis of statistical analysis we found that there were no influences on the expression of CD44v6 by the TAN II A and the concentration of oxygen. The TANⅡA can not regulate the expression of HIF-1α.Reconstruct experiment of the invasion of basement membrane and measure the adhesion of tumor cell and basement membrance:the invasion of PG cell improved in hypoxic state. And the hypoxic state is good for PG cell's metastasis. however the TANⅡA can not regulate the invasion of PG cell.