摘要
目的miR-320a作为微小RNA(micro RNA,miRNA)类型之一,在多种恶性肿瘤组织中表达异常,与肿瘤发生、进展及预后关系密切。本研究分析卵巢上皮性癌组织中miR-320a表达,并探讨上调人卵巢癌SKOV3细胞中miR-320a表达对癌细胞增殖和侵袭力的衫响。方法选取2015-02-02-2018-03-21在湖北省中医院妇产科收治的94例患者的上皮性卵巢癌组织,选取同期42例正帯卵巢组织标本。实时荧光定量PCR检測上皮性卵巢癌和正常卵巢组织中miR-320a表达。培养SKOV3细胞并分为miR-320a模拟组、阴性对照组和空白组。实时荧光定量PCR检测细胞中miR-320a表达,MTT法和克隆形成实验检测细胞增殖力,Transwell法检测细胞侵袭力,蛋白质印迹法检测细胞中E-cadherin、N-cadherin和Vimentin蛋白表达。结果卵巢癌组织中miR-320a相对表达量为0.51±0.09,低于正常卵巢组织的1.00±0.05,差异有统计学意义,t=32.448,P<0.001。上皮性卵巢癌组织中miR-320a表达量在FIGO分期(t=4.450,P<0.001)、分化程度(t=2.348,P=0.021)和有无淋巴结转移(t=3.728,P<0.001)方面比较,差异有统计学意义。miR-320a模拟组细胞中miR-320a相对表达量高于阴性对照组和空白组,差异有统计学意义,F=223.704,P<0.001。MTT实验结果显示,3组细胞活力均值间差异有统计学意义,F组间=7 259.713,P<0.001;miR-320a模拟组抑制细胞活性高于其他2组。克隆形成实验结果显示,miR-320a模拟组细胞克隆数低于阴性对照组和空白组,差异有统计学意义,F=975.124,P<0.001;miR-320a模拟组侵袭细胞数低于阴性对照组和空白组,差异有统计学意义,F=124.195,P<0.001;miR-320a模拟组细胞中E-cadherin蛋白相对表达量高于阴性对照组和空白组,差异有统计学意义,F=152.673,P<0.001;而N-cadherin和Vimentin蛋白相对表达量低于阴性对照组和空白组,差异有统计学意义,F值分别为55.251,62.054,均P<0.001。结论miR-320a在上皮性卵巢癌组织中呈低表达,且与肿瘤发生及恶性化进展有关。上调SKOV3细胞中miR-320a表达可抑制细胞增殖及细胞侵袭力,其机制可能与抑制上皮-间质转化有关。
OBJECTIVE miR-320a is one of the miRNA types.Previous studies have shown that it is abnormally expressed in many malignant tumor tissues,and is closely related to tumorigenesis,progression and prognosis.The purpose of this study was to analyze the expression of miR-320a in epithelial ovarian cancer,and to explore the effect of up-regulate the expression of miR-320a on proliferation and invasiveness of human ovarian cancer SKOV3 cells.METHODS A total of 94 cases of patients with epithelial ovarian cancer underwent surgery in Hubei University of Traditional Chinese Medicine were selected from February 2015 to March 2018.In the same period,42 cases of normal ovarian tissue specimens were selected.The expressions of miR-320a in epithelial ovarian cancer and normal ovarian tissues were detected by realtime fluorescent quantitative PCR.SKOV3 cells were cultured and divided into miR-320a mimic group,negative control group and blank group.Theexpression of miR-320a in cells was detected by real-time fluorescent quantitative PCR.Cell proliferation was detected by MTT assay and colony formation assay.Cell invasiveness was detected by using Transwell assay.The expressions of E-cadherin,N-cadherin and Vimentin proteins were detected by Western blot.RESULTS The relative expression level of miR-320a was 0.51±0.09 in ovarian cancer tissues,which was lower than that in normal ovarian tissues,which was 1.00±0.05,the difference was statistically significant(t=32.448,P<0.001).Comparison on the expression levels of miR-320a in epithelial ovarian cancer tissues in FIGO stage(t=4.450,P<0.001),degree of differentiation(t=2.348,P=0.021)and lymph node metastasis(t=3.728,P<0.001),the differences were statistically significant.The relative expression level of miR-320a in miR-320a mimic group was higher than that in negative control group and blank group,the difference was statistically significant(F=223.704,P<0.001).The results of MTT assay showed that the differences of cell viability among the three groups were statistically significant(Fgroup=7 259.713,P<0.001).The cell viability in the miR-320a mimic group was higher than in the other two groups.The results of colony formation assay showed that the number of cell clones in miR-320a mimic group was lower than that in negative control group and blank group,the difference was statistically significant(F=975.124,P<0.001).The number of invasive cells in miR-320a mimic group was lower than that in negative control group and blank group,the difference was statistically significant(F=124.195,P<0.001).The relative expression level of E-cadherin prot&n in miR-320a mimic group was higher than that in negative control group and blank group,the difference was statistically significant(F=152.673,P<0.001),while the relative expression levels of N-cadherin and Vimentin proteins were lower than those in negative control group and blank group,the differences were statistically significant(F values were 55.251 and 62.054,P<0.001).CONCLUSIONS miR-320a is lowly expressed in epithelial ovarian cancer tissues,and is related to tumorigenesis and malignant progression.Up-regulation expression of miR-320a in SKOV3 cells could inhibit cell proliferation and cell invasion,and its mechanism may be related to inhibition of epithelial-mesenchymal transition.
作者
余俊
李晓兰
张青冬
YU Jun;LI Xiao-lan;ZHANG Qing-dong(Department of Obstetrics and Gynecology,Hubei University of Traditional Chinese Medicine,Traditional Chinese Medicine Hospital of Hubei,Wuhan 430061,P.R.China;Department of Obstetrics and Gynecology,Second People Hospital of Yichang,Yichang 443000,P.R.China;Department of Obstetrics and Gynecology,Huangshi Central Hospital Affiliated Hospital of Hubei Polytechnic University,Edong Healthcare Group,Huangshi 435000,P.R.China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2019年第14期1026-1030,1035,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
湖北省自然科学基金(2015CFB574)
