| The incidence of primary lung cancer ranks first in the tumor, the various means of comprehensive treatment has a positive effect on the prevention and treatment of cancer, but the result is not satisfied, so we are facing very urgent situation. The reasons for this situation including:increased cancer incidence rate, fewer cases of early detection, significant side effects of drugs, enhanced tumor resistance, so early detection of the disease to control incidence rates, effective treatment to reduce mortality has become the goal of the study. Tumor angiogenesis plays an important role in the occurrence, development, invasion, metastasis and recurrence of cancer. Therefore, anti-angiogenic therapy combined with cytotoxic drugs may increase the effect of the treatment of cancer. Pro-angiogenic factor and anti-angiogenic factor are the most important in the regulation of angiogenesis generation, It is crucial to maintain homeostasis.Hypoxia-inducible factor 1α( HIF-1α) and Basic fibroblast growth factorb,(b-FGF) are the most important pro-angiogenic factors. Vasohibin-1,VASH-1 is the only negative feedback control angiogenesis factors that is discovered. Relationship among lung cancer has not been reported.Antiangiogenic therapy combined with cytotoxic drug provided the new direction to the treatment of cancer. Traditional chinese medicine has become an anti-tumor adjuvant, TMP, as one of them, has some anti-tumor effect. The experiment through dynamic observation of weight and tumor growth in different groups(control group,cisplatin groups,TMP group,combination group) of Lewis lung cancer C57BL/6 mice model, the change of the tumor cells and tumor vascular morphology, quality of tumor,and changes in the expression of VASH-1, HIF-lα, b-FGF, can explain thatTMP combined with cisplatin may improve the therapeutic effect, and can describe the possible mechanism, so as to provide an experimental basis for TMP widely used in clinical.The process of experimental is as follows.Cultured Lewis lung carcinoma cells and injected the right armpit of mice. Two weeks later,tumor diameter was about 8 mm. The tumor-bearing mice were randomly divided into four groups namely: control group, TMP group, cisplatin group, combination group, 12 rats in each group, cisplatin at a dose of 2 mg·kg-1, ligustrazine at a dose of 100 mg· kg-1, combination group(the doses as above), all mice were sacrificed after treatment for two weeks and these tumors were obtained, weighed, and the tumor inhibitory was calculated. The change of cell and vascular structure was observed by HE,TEM and the change of HIF-1α,b-FGF and VASH-1 were determined by immunohistochemistry and Western Blot after treatment. Take the saline group as standard: Diet, activity, mental status of TMP group did not change significantly, while changes in the cisplatin group is reducing diet and activity, poor mental state and dull hair; the mice crouched together into groups; The situation in combined group is between the two situations.The change of tumor volume was as follows: TMP group>cisplatin group >combination group; The changes of body weight and tumor weight had the same trend with tumor volume.The result of the inhibition rate showed :Compared with the control group, the tumor inhibition rate in experimental groups was significantly increased(P<0.05). It was the highest in combination group(50.24%)and cisplatin group was better than the TMP group(P<0.05). Microscopy and transmission electron microscopy showed: cell size, cell organelles, chromosomes, vascular structures in those groups have changed compared with those in saline group. The change of TMP group is significantly weaker than cisplatin,combination group is the largest, observing apoptotic bodies. Immature blood vessels can be seen in the saline group while other groups was relatively normal and mature. The expression of HIF-1α, b-FGF andVASH-1 showed by Immunohistochemistry: These in experimental group were decreased obviously compared with control group(P<0.05), and they were the lowest in the combined group and the highest in control group.The result showed by Western Blot Western blot assay:They had the same trend. Correlation analysis showed: HIF-1α is positively correlated with b-FGF(r =0.610, P=0.035), b-FGF is positively correlated with VASH-1(r=0.591, P=0.043), HIF-1α is uncorrelated with VASH-1(r= 0.171, P =0.549).By observing general condition of the mice,weight,volume changes,the expression of angiogenesis-related factor,tumor cell and the comparison of mean tumor weightand inhibition rate,it is proved that TMP can not only anti-angiogenesis,but also enhance the effect of cisplatin in the treatment of tumors.The mechanism of TMP combined with cisplatin may be:By TMP inhibiting the expression of HIF-1α,downward the transcription and translation of b-FGF gene,resulted in the induction of b-FGF reduced ability,ultimately reduced expression of the VASH-1,maintained and regulated dynamic equilibrium between the angiogenic factors and promoted normalization of blood vessels.Then,cisplatin reaches the site of tumor,mitigating damage to normal cells,reducing side effects.VASH-1 inhibits angiogenesis,and its successful development of biological agents may play a positive role in angiogenesis therapy.ACRONYM NAME IN ENGLISH中文全称DMEM dulbecco’s modified eagle medium Eagle培养基NSCLC non-small cell lung cancer非小细胞肺癌HIF-1αhypoxia-inducible factor 1 alpha缺氧诱导因子1αVASH Vasohibin血管生成抑制蛋白DAB 3,3-Diaminobenzidine 3,3-二氨基联苯胺IHC immunohistochemistry免疫组织化学法TMP tetramehtylpyrazine川芎嗪PBS phosphate buffer saline硫酸盐缓冲液b-FGF Basic fibroblast growth factorb,碱性成纤维细胞生长因子COX-2 cyclo-oxygen-ase环氧化酶2IOD integral optical density平均光密度VEGF Vascular endothelial growth factor血管内皮生长因子... |
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