微小RNA-384通过下调多效生长因子表达抑制胰腺癌细胞增殖及浸润  

MicroRNA-384 inhibits proliferation and invasion of pancreatic cancer cells by down-regulating pleiotropic growth factor expression

作　　者：符常波[1] 聂磊[1] 殷涛[1] 吴东德[1] Fu Changbo;Nie Lei;Yin Tao;Wu Dongde(Department of Hepatopancreatobiliary Surgery,Cancer Hospital of Hubei Province,Wuhan 430079,China)

机构地区：[1]湖北省肿瘤医院肝胆胰科,武汉430079

出　　处：《中华实验外科杂志》2019年第6期1048-1051,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察微小RNA(miRNA,miR)-384在胰腺癌组织及癌旁组织、胰腺癌细胞株中的表达及其与胰腺癌临床病理特殊的关系及其对癌细胞株增殖、迁移等的影响,探讨其作用机制。方法查询生物信息预测miR-384可能靶向结合多效生长因子(PTN)的mRNA,采用反转录-聚合酶链反应(RT-PCR)技术检测2016年3月至2018年12月湖北省肿瘤医院67例胰腺癌组织标本或穿刺标本中癌及癌旁组织中miR-384及PTN的表达,用携带miR-384的腺病毒转染胰腺癌细胞株PANC-1/MIAPaCa-2,PT-PCR技术检测转染前后miR-384、PTNmRNA表达量,蛋白质印迹法(Westernblot)检测转染前后PTN蛋白变化,通过噻唑蓝(MTT)比色法、Transwell小室检测转染前后PANC-1/MIAPaCa-2增殖、迁移及侵袭能力变化。应用SPSS19.0统计软件和GraphPadPrism8.0进行数据处理和统计分析,数据以均值±标准差(Mean±SD)表示。miR-384在癌组织和癌旁组织间表达量的比较采用配对样本的t检验。结果miR-384在胰腺癌组织及癌旁组织中相对表达量分别为0.0037±0.0009、0.0065±0.0022,差异有统计学意义(t=6.177,P<0.01),并与神经浸润及淋巴结转移明显相关(χ^2=4.750、13.400,P<0.05)。将胰腺癌组织分为miR-384低表达组及高表达组,PTNmRNA相对表达量分别为0.073±0.028、0.045±0.001,差异有统计学意义(t=2.082,P<0.05)。转染腺病毒后,与阴性对照组比较,PANC-1细胞中miR-384表达提高11.69倍(t=28.600,P<0.01),PTNmRNA及PTN蛋白则分别降至19.02%(t=31.200,P<0.01)、38.00%(t=16.200,P<0.01),MIAPaCa-2细胞中miR-384表达提高14.95倍(t=21.300,P<0.01),PTNmRNA及PTN蛋白分别降至15.02%(t=13.300,P<0.01)、13.74%(t=13.500,P<0.01),差异有统计学意义。两种细胞株增殖、迁移、侵袭能力均下降,而重组人PTN蛋白处理可逆转此情况。结论miR-384在胰腺癌中具有明显抑制肿瘤作用,其机制可能是通过靶向结合PTNmRNA下调PTN表达实现。Objective To study the expression of microRNA (miRNA, miR)-384 in pancreatic cancer, para-carcinoma tissue and pancreatic cancer cell lines as well as the relationship between miR-384 and clinical pathology of pancreatic cancer, and the effects of miR-384 on proliferation and migration of pancreatic cancer cell lines. Methods The expression of miR-384 and pleiotropic growth factor (PTN) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in 67 pancreatic cancer tissues and corresponding adjacent tissues obtained by resection or aspiration biopsy. The expression of miR-384 and PTN mRNA and PTN protein in pancreatic cancer cell line PANC-1/MIAPaCa-2 was detected by RT-PCR and Western blotting before and after the transfection of adenovirus loaded with miR-384. The influence of the infection to the proliferation, migration and invasion of PANC-1/MIAPaCa-2 were detected by methyl thiazol tetrazolium (MTT) Assay and Transwell chamber. Results The relative expression of miR-384 was 0.003 7±0.000 9 and 0.006 5±0.002 2 in pancreatic cancer tissues and adjacent tissues respectively (t=6.177, P<0.01), and was associated with nerve invasion (χ^2=4.750, P<0.05) and lymph node metastasis (χ^2=13.400, P<0.01). While the pancreatic carcinoma tissues were divided into miR-384 low-expression group and high-expression group, the relative expression of PTN mRNA was 0.073±0.028 and 0.045±0.001, respectively, the differences of which were statistically significant (t=2.082, P<0.05). Compared with the negative control group, the adenovirus transfection group reveals that the expression of miR-384 in PANC-1 cells was increased 11.69 times (t=28.600, P<0.01), while the expression of PTN mRNA and PTN protein were decreased to 19.02%(t=31.200, P<0.01) and 38.00%(t=16.200, P<0.01) respectively, and the expression of miR-384 was increased 14.95 (t=21.300, P<0.01) times in MIAPaCa-2 cells while PTN mRNA and PTN protein decreased to 15.02%(t=13.300, P<0.01) and 13.74%(t=13.500, P<0.01) respectively. The proliferatio

关 键 词：胰腺癌 多效生长因子 侵袭转移 RNA干扰 微小RNA-384 

分 类 号：R735.9[医药卫生—肿瘤]