| Esophageal cancer is one of the most common malignant tumor of digestive tract. According to the report of world esophageal cancer incidence and mortality, the incidence cases number of esophageal cancer was about 481,600, and the number of death cases was about 406,500 in 2008 in the world[1]. Both morbidity and mortality of EC varied greatly by regions, nation, race, gender and over time. In particular, China is one of countries with high incidence and mortality rates of EC area[2]. In general, the etiology is still unclear although some theories are proposed recently. Patients with digestive tract cancer usually have abnormal expression of mi RNA[3]. Konishi [4]studied 106 patients with EC and 54 healthy volunteers as controls and found that expression level of mi RNA-18 a in serum for EC was significantly higher than one of control patients and close to 16 times. In addition, its level in EC tissues was significantly higher than that of surrounding normal tissues. Its level in serum of EC patients was significantly lower after surgical resection of tumor. Many recent studies indicated that mi RNA-1 expression level were associated with the proliferation of esophageal squamous cell carcinoma(ESCC). However, its effect and related mechanism are still not well-studied. That is the focus of this research. In short, mi RNA is a length of about 20-22 nucleotides characterized with non coding small molecule RNA[4]. Its relationship with stages of tumor and potentials of mi RNA in early diagnosis and treatments of EC has been reported [13].Objective: In order to investigate the mechanism of mi RNA-1 in ESCC, we transfected mimics-mi R-1, mimics-mi R-1control, inhibitor-mi R-1 and nhibitor-mi R-1 control into ESCC cell line KYSE510 and compared their effects on cell proliferation, migration and the protein expression of target gene.1 The experiment designs experimental group and control group, experimental group was mimics-mi R-1 group which mi R-1 over-expressed in KYSE510 cell line.The cell line was transfected with mimics which contained the element of mi R-1 simulation material. And its control group was mimics-mi R-1 control group.The cell line was transfected with simulation reagent of mimics which other components was same. Another experimental group was inhibitor-mi R-1 group. The expression of mi R-1 was inhibited in KYSE510 cell line.This cell line was transfected with Inhibitor which contained the element of mi R-1 inhibitor. Its control group was mimics-mi R-1 control group.The cell line was transfected with simulation reagent of inhibitor which other components was same.2 MTT cell proliferation test was used to observation the different cell proliferation which was affected by mi RNA-1 over-expression and inhibition in KYSE 510 cell line.3 Transwell test was used to observe the different cell migration and invasion which were affected by mi RNA-1 over-expression and silencing in KYSE 510 cell line.4 The expression of LASP1 and TAGLN2 were determined with western blot.Results: 1 Methylthiazolyldiphenyl-tetrazolium bromide test 1.1 The OD values of 24 h, 48 h, 72 h in the experimental group which mi R-1 was over expression and was transfected by mimics-mi R-1 were 0.894±0.023, 1.050±0.013, 1.232±0.017. And its control group OD values were 0.963±0.022,1.163 ± 0.030, 1.377 ± 0.024. Experimental group OD values were lower than control group. And the experiment group OD values which mi R-1 was inhibited and was transfected with inhibitor-mi R-1 at 24 h, 48 h, 72 h were 0.963±0.022, 1.163±0.030, 1.377±0.024, respectively. The control group OD values were 0.968 ± 0.067, 1.209 ± 0.027, 1.410 ± 0.023. Experimental group OD values were higher than control group. 1.2 Over expression of mi R-1 in ESCC cell line KYSE510 inhibitedMethods: KYSE510 cell line proliferation. The inhibition of mi R-1 expression in ESCC cell line KYSE510 promoted KYSE510 proliferation. 2 Transwell chamber experiment 2.1 The experimental group which was transfected with mimics cell number was 112±24 in migration experiments. And the cell number in control group which was transfected with simulation reagent of mimics was 230±32. The low expression of mi RNA-1 experimental group cell number was 181±29 in migration experiments and the reagent is inhibitor-mi R-1. The cell number of the control group was 130±25. 2.2 The cell number in experimental group which mi R-1 was over expression was 125±31 in invasion assay. The cell number in its control group was 204±42. In Invasion assay the experimental group cell number which mi R-1 was inhibited was 235±36. The cell number in its control group was 116±25. 2.3 The experiment results show that when mi R-1 over express, ESCC cell line KYSE510 was inhibited in migration and invasion. When mi R-1 expression was inhibited in ESCC cell line KYSE510, the ability of migration and invasion was enhanced. 3 Western Blot the experimental result 3.1 This experimental group mi R-1 was over expression and its relative expression of LASP1 was 0.498±0.020, which was lower than that its control group 0.642±0.070. And the relative expression of LASP1 in experimental group which mi R-1 was inhibited was 0.626±0.050, and the expression value was higher than its control group 0.526±0.030. 3.2 The relative expression of TAGLN2 in mi R-1 over expression experimental group was 0.370±0.020, which was lower than its control group 0.426±0.030. And the relative expression of TAGLN2 in experimental group which was transfected with inhibitor-mi R-1 was 0.541±0.050 and it was higher than that its control group0.421±0.051. 3.3 When mi R-1 over expressed, the level of relative expression of LASP1 and TAGLN2 was lower and when mi R-1 expression was inhibited in ESCC cell line KYSE510, the level of relative expression of LASP1 and TAGLN2 was higher than control group.Conclusions:1 Over expression of mi R-1 in cells can inhibit the esophageal squamous cell carcinoma cell(KYSE510) proliferation and weaken the migration and invasion. Inhibition of expression levels of mi R-1 in cells can enhance the esophageal squamous cell carcinoma cell(KYSE510) proliferation and enhance the ability of migration and invasion.2 The level of mi R-1 expression regulates the expression of LASP1, TAGLN, which may be the regulation mechanism. |
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