Expression Of Substance P In Endometrial Adenocarcinoma And Its Significance

Objective: As one of the three most common malignant tumor of female reproductive system,the morbidity and mortality of Endometrial Carcinoma(EC)in the global women were increasing.According to incomplete statistics,EC accounts for about 20%-30% in the female genital system neoplasms which seriously threaten the physical and mental health of the majority of women.The most common pathological type was adenocarcinoma.With the increase of estrogen and progesterone in the treatment of non regular replacement therapy since the 20 th century,it shows a significant rise in incidence of EC in women in China.The occurrence and development of endometrial carcinoma is a complex pathological process,and its etiology and pathogenesis have not yet been explained clearly.With the development of molecular biology and immunology,more and more researchers are focused on many kinds of molecules or signaling pathways.Substance P(SP)has attracted much attention because of its wide distribution.Substance P is a small molecular polypeptide composed of 11 amino acid residues with a molecular weight of 1340.It is a number of tachykinin family which was encoded by TAC-1 gene.Substance P is widely expressed in central nervous system and peripheral tissues in our body,also with a variety of biological activity.Substance P can be involved in the regulation of the function of immune,and enhance the repair of nerve tissue after trauma.In addition,substance P has a proliferation promoting effect on a variety of cells.When the body is subjected to traumatic stimulation,substance P can induce the proliferation of vascular endothelial cells and fibroblasts.In recent years,with the in-depth study of scholars at home and abroad,the role of substance P in tumor is gradually being known.studies have found that the expression of substance P has increased significantly in a variety of tumors such as breast cancer,stomach cancer,pancreatic cancer,Laryngeal cancer,lung cancer,melanoma etc.However,the expression of substance P in endometrial cancer is rarely mentioned till now.Therefore,this study aims to use Quantitative Real-time Polymerase Chain Reaction,Western Blotting to measure the expression of substance P in Endometrial adenocarcinoma tissue and Ishikawa cell strains and to use MTT and Transwell experiments to explore the relationship of the expression intensity of substance P with endometrial cancer cell proliferation and movement.In conclude,this study aims to discuss the role and meaning of substance P in the occurrence and development of disease.Methods:1 The expression of substance P(mRNA)in Endometrial adenocarcinoma tissue and Ishikawa cells was detected by qPCR technique: Detect the expression of mRNA of substance P in those groups,by using Endometrial adenocarcinoma tissue and Ishikawa cells as the experimental groups and normal endometrial tissue as the control group.2 Detection of protein content of substance P in Endometrial adenocarcinoma tissue and Ishikawa cells by Western Blotting method: Detect the expression of protein of substance P in those groups,by using Endometrial adenocarcinoma tissue and Ishikawa cells as the experimental groups and normal endometrial tissue as the control group.3 Vitro proliferation of different concentrations of substance P on Ishikawa cells after 48 h has been detected by MTT: different concentrations of substance P(10-9? 10-8? 10-7?10-6mol/l)have been respectively added to Ishikawa cells which were in logarithmic phase.Placed in a constant temperature incubator(5% CO2,37?)for 48 h,then,detected the Vitro proliferation by MTT.4 Method of Transwell was used to test the changes of invasive ability of Ishikawa cells within different concentrations of substance P: different concentrations of substance P were added to Ishikawa cells,after coating the Transwell small room by Matrigel,join groups of cells drop by drop,placed in a constant temperature incubator(5% CO2,37?)for 24 h,immobilization and dyeing,count numbers of groups of cells in a falling hole.5 Statistical methods: statistical software SPSS 13.0 were used to analyzed,and the experiment data meet normal distribution,the measurement data to mean ± standard deviation,t test and single factor analysis of variance comparing differences between groups,?=0.05 for the inspection level,when p <0.05 there was statistically significant.Result:1 The qPCR technology detects the expression conditions of substance P(mRNA)in Endometrial adenocarcinoma tissue;Ishikawa cells and normal endometrial tissue: The expression quantity of P material in Endometrial adenocarcinoma tissue was 2.62±0.13,in Ishikawa cells was 3.65±0.28,in normal tissue was 1.02±0.02,and the expression of substance P in endometrium gland cancer tissue and cells were obviously higher than normal tissue,their difference owns the statistical significance(P<0.001).2 The protein amount of substance P expressed in Endometrial adenocarcinoma tissue;Ishikawa cells and normal endometrial tissue detected with the method of Western Blotting: The results shown that the expression of substance P protein in Endometrial adenocarcinoma tissue;Ishikawa cells line and normal tissue were respectively 1.06±0.09;1.20±0.11 and 0.82±0.06.The expression of substance P in adenocarcinoma of endometrium was obviously higher than normal tissue,their difference owns the statistical significance(P<0.001).3 The rate of increase of different concentration P material(10-9?10-8?10-7?10-6mol/l)acting on adenocarcinoma of endometrium Ishikawa cell detected with MTT method for forty-eight hours was respectively : 8.933±0.615?14.667±1.195?26.523±3.374?20.983±1.911.The result shown: when the substance P concentration was from 10-9mol/l to 10-7mol/l,and cultivated together with Ishikawa cells for forty-eight hours,with the addition of substance P concentration,the cell reproductive capacity gradually increase,while when substance P added to 10-6mol/l,the cell reproductive capacity decline to some extent.The difference among each line was obvious(P<0.01),which had the statistical significance.4 The invasive cell population of Ishikawa cell detected by Matrigel-Transwell invasion assay after the treatment of different concentration substance P: In the experiment of invasion,the substance P concentration chooses the range from 10-9mol/l to 10-7mol/l,when the substance P was cultivated with Ishikawa cells for forth-eight hours,the quantity of tumor cells across the matrigel was respectively: 26±5.612?37.6±7.603?49.6±9.737,the invasion capability gradually increase with the addition of Substance P concentration.The difference among each lines was obviously(P<0.01),which had the statistical significance.Conclusion:1 The substance P was significantly over expression in Endometrial adenocarcinoma than that in the normal endometrium.It suggests that substance P may be involved in the occurrence and development of endometrial cancer.It is hoped to provide a new serological marker for the diagnosis of endometrial adenocarcinoma.2 The substance P can significantly strengthen the proliferation of endometrial adenocarcinoma cells.It shows that the substance P is related to the occurrence of endometrial cancer,which is helpful to explain the pathogenesis of endometrial adenocarcinoma.3 The substance P can significantly enhance the invasive ability of endometrial adenocarcinoma cells.In the experimental concentration range,concentration is higher in promoting the invasion ability is stronger.It suggests that substance P may play an important role in the malignant transformation of endometrial cancer which is helpful to judge the prognosis of endometrial adenocarcinoma.