| In recent years, the incidence rate of malignant tumor is rising. Duringall the malignant tumors, lung cancer has become the primary cause of death. Tough people have made great progress in diagnosis and treatment of lung cancer, total five years suevival rate of patients has not improved which is just10%The reason for this phenomenon is that malignant has two important bilolgical characteristics which is invasion and metastasis. Invasion and metastasis are two complex processes that include a plurality of link and steps in which cell adhesion, matrix degradation and cell movement play an important role. How to control the invasion and metastasis of malignant tumors has become an important topic in the area of tumor in recent years. Seeking and exploring effective methods and drugs to prohibite invasion and metastasis of tumor is significant to improve the patients suevival rate and reduce postoperative recueuence rate.Most of malignant tumor patient have blood stasis ayndrome,which can be proved by the clinical manifestations, abnormal blood rheology, microcirculation obstacle and blood coagulation state. Blood stasis syndrome is not only a objective existence pathological of patients with malignant tumor but also a favorable metastasis factor. Tumor cells can induce platelet increased and aggregation through such mechanisms as cell adhesion. And platelet can secret coagulation factor which lead to the blood high coagulation state,causing the formation of microthrombus which including cancer cells.The formation of microthrombus contribute to the matastasis of tumor. At the meantime, cancer cells have unlimited proliferative ability, which inevitably leads to a hypoxic condition as a result of the rapid proliferation of tumor tissue. Thus, hypoxic survival is one of the biological characteristics of malignant tumor, and is also considered as a malignant tumor metastasis factor.Application of blood activating drugs to cure tumor is a commonly used clinical treatment, however, reserches still have different opinions about the effect to the metastasis of tumor which bought by the blood activating drugs. Some research consider that blood activating drugs can inhibit the invasion and metastasis of tumor,other research think that blood activating drugs contribute to the invasion and metastasis of tumor. Also, there ara some research believe that blood activating drugs can not only promote but also inhibit the invasion and metastasis of tumor. The above discrepancy may has a close relationship with the various blood activating drugs, the vary patient’s symptoms and different drug combination. According to previous research, we hypothesize that blood activating drugs may in some process contribute the tumor metastasis, and in some other process inhibit tumor metastasis.Based on the above viewpoints, in study we selected the effective components of blood activating deugs Chuanxiong and Salvia miltiorrhiza which named TMP and tanshinone Ⅱ A.We use TMP and tanshinone Ⅱ A to intervene the human high metastatic large cell lung cancer cell PG, and then we inoculate on nude mice, causing occurrence of spontaneous metastasis. Through the detection of in primary tumor weight and the tumor inhibition rate of in primary tumor, expression of E-cad, number of lung metastases and inhibition of lung metastasis rate, lung CD44v6expression, expression of serum P-select, liver VEGF expression, we study the effect of transfer ability on PG cell which bought by the blood activating drugs, and thy to analyze the mechanism from the point of cell adhesion and angiogenesis.Research methodCulture PG cells in normoxia (5%CO2,95%air) and oxygen (5%02,5%C02,90%N2) environment, and use TMP and tanshinone Ⅱ A to intervene PG celld respectively. At the same time, set up a blank control group, negative control group, DMSO control group and positive control group DDP. Every group of cells are intervened by different drugs and then inoculated in the right subaxillary of nude mice. Kill the nude mice after six weeks later and then the detect relevant indicators through immunohistochemistry and Elisa method.Research resultsHumor inhibition rate of in primary tumorIn oxygen environment, the difference between TMP group and control group is statistically significant (P=0.016, P<0.05). The difference between tanshinone Ⅱ A group and control group is statistically significant (P=0.016, P<0.05). In hypoxic environment, compared with the control group, both the TMP and tanshinone Ⅱ A group are notable statistically significant (P<0.0001). The different between TMP and tanshinone Ⅱ A is not statistically significant (P>0.1). The different between hypoxia control and normoxia control group is not statistical significant. The above results suggest that both TMP and tanshinone Ⅱ A in hypoxia and normoxia environment have certain prohibitive effect on the development of tumor. Compared to the hypoxia environment, the normosxia environment has certain prohibitive effect on the development of tumor., but the differende is not significant.2E-Cad expression of primary tumorIn normoxia environment, the difference of E-Cad expression between the TMP group and control group is statistically(p=0.012, p<0.01). The difference of E-Cad expression between tanshinone Ⅱ A group and control is notable statistically significant(p<0.001).In hyperxia environment, both the TMP group and tanshinone Ⅱ A group have significant differences with the control group(P<0.0001).The difference between TMP and tanshinone Ⅱ A group is not statistically significant (p>0.1).The difference between normoxia control and hyperxia control group is not statistically significant (P>0.1). The above result suggest that both TMP and tanshinone Ⅱ A can upregulate the expression of E-Cad. Different oxygen environment have no influence on E-Cad expression.3Inhibition of lung metastasis rateIn normoxia environment, the difference of inhibition of lung metastasis rate between TMP and control group is statistically significant (p=0.085, p<0.1) The difference of inhibition of lung metastasis rate between tanshinone Ⅱ A group and control group is statistically significant(p=0.085, p<0.1). In hypoxic environment, the difference of inhibition of lung metastasis rate between TMP and control group is statistically significant(p=0.028, p<0.05). The difference of inhibition of lung metastasis rate between tanshinone Ⅱ A group and control group is statistically significant (p=0.025, p<0.05). The different between TMP and tanshinone Ⅱ A is not statistically significant (P>0.1). The different between hypoxia control and normoxia control group is no statistical significant. The above results suggest that both TMP and tanshinone ⅡA can at a certain prohibit the lung metastasis rate,there have no, however,dierence between the normoxia and hyperxia environment.4Expression of lung CD44v6In normoxia environment, the difference of CD44v6expression of lung tissue between TMP and control group is statistically significant (p=0.004, p<0.05) The difference of inhibition of CD44v6expression of lung tissue between tanshinone II A group and control group is statistically significant (p=0.003, p<0.05). In hypoxic environment, the difference of inhibition of CD44v6expression of lung between TMP and control group is statistically significant ’(p=0.002, p<0.05). The difference of CD44v6expression of lung between tanshinoneⅡ A group and control group is statistically significant (p=0.002, p<0.05). The different between TMP and tanshinone Ⅱ A is not statistically significant (P>0.1). The different between hypoxia control and normoxia control group is statistical significant (p=0.028, p<0.05). The above results suggest that both TMP and tanshinone ⅡA group can prohibit the expression of CD44v6in lung tissue. Besides, the hyperxia can increase the espression of CD44v6in lung cancer.5Expression of serum P-selectionIn normoxia environment, the difference of expression of serum P-selection between TMP and control group is statistically significant (p=0.021, p<0.05) The difference of inhibition of expression of serum P-selection between tanshinone Ⅱ A group and control group is statistically significant (p=0.01, p<0.05). In hypoxic environment, the difference of inhibition of expression of serum P-selection between TMP and control group is statistically significant (p=0.016, p<0.05). The difference of CD44v6expression of lung between tanshinone Ⅱ A group and control group is statistically significant (p=0.072, p<0.1). The different between TMP and tanshinone Ⅱ A is not statistically significant (P>0.1). The different between hypoxia control and normoxia control group is not statistical significant (p>0.1).The above results suggest that both TMP and tanshinone ⅡA group can prohibit the expression of P-selection in serum. However, there are mo difference between the normoxia and hyperxia environment.6Expression of liver VEGFIn normoxia environment, the difference of expression of liver VEGF between TMP and control group is statistically significant (p=0.01, p<0.05). The difference of inhibition of expression of liver VEGF between tanshinone Ⅱ A group and control group is statistically significant (p=0.011, p<0.05). In hypoxic environment, the difference of expression of liver VEGF between TMP and control group is statistically significant (p=0.024, p<0.05). The difference of liver VEGF betweentanshinone Ⅱ A group and control group is statistically significant (p=0.034, p<0.05). The different between TMP and tanshinone Ⅱ A is not statistically significant (P>0.1). The different between hypoxia control and normoxia control group is not statistical significant (p>0.1).The above results suggest that both TMP and tanshinone ⅡA group can prohibit the expression of liver VEGF. However, there are mo difference between the normoxia and hyperxia environment. |
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