摘要
目的探讨肺腺癌组织丝氨酸蛋白酶抑制因子Kazal 1型(SPINK1)表达与生存时间的关系及其对肺腺癌细胞生长的影响。方法用Real-time PCR法、免疫组织化学法检测285例肺腺癌组织和癌旁组织SPINK1的表达,并分析其表达水平与临床病理特征及预后的关系。利用SPINK1慢病毒表达载体及小干扰RNA(si RNA),处理人肺腺癌细胞系,光学显微镜下观察绿色荧光蛋白(GFP)表达情况,ELISA或Real-time PCR检测SPINK1表达水平,细胞计数和集落形成法检测肺腺癌细胞生长情况。结果肺腺癌组织SPINK1 mRNA和蛋白表达均上调(均P<0. 0001),SPINK1高表达组患者总生存时间短于低表达组(50. 0个月vs 64. 5个月,P<0. 001)。SPINK1过表达组,肺腺癌细胞上清液SPINK1蛋白高表达(P <0. 05),增殖与克隆形成能力均上调(P <0. 05);SPINK1敲减组,肺腺癌细胞SPINK1 mRNA表达下调,增殖能力下调(P<0. 05)。结论肺腺癌组织SPINK1高表达提示患者预后不良; SPINK1促进人肺腺癌细胞的增殖。
Objective To explore the relationship between serine protease inhibitor Kazal type 1( SPINK1 expression and survival time of lung adenocarcinoma( LAC) and its effect on the growth of LAC cells. Methods The SPINK1 expressions in 285 cases of lung adenocarcinoma tissues and adjacent tissues were detected by Real-time PCR and immunohistochemical staining. The relationship between SPINK1 expression and clinicopathological features and prognosis of lung adenocarcinoma was analyzed. Human LAC cells were treated with lentivirus vector with SPINK1 or SPINK1 siRNA.Green fluorescent protein( GFP) expression was observed under microscope. SPINK1 expression was detected by enzymelinked immunosorbent assay( ELISA) or Real-time PCR. The growth of LAC cells was evaluated by cell count and colony formation assay. Results The levels of SPINK1 mRNA and protein level in lung adenocarcinoma were up-regulated( P<0. 0001). The overall survival time in the group of SPINK1 high expression was shorter than that of SPINK1 low expression( 50. 0 months vs 64. 5 months,P< 0. 001). In SPINK1 overexpression group,SPINK1 protein was up-regulated in the cultures of LAC cells( P < 0. 05),and the ability of proliferation and colony formation was up-regulated( P < 0. 05). In SPINK1 knockdown group,SPINK1 mRNA was down-regulated in LAC cells( P < 0. 05),and the ability of proliferation was decreased( P < 0. 05). Conclusion High expression of SPINK1 in lung adenocarcinoma suggests poor prognosis.SPINK1 promotes the proliferation of LAC cells.
作者
陆畅畅
黄燕燕
续力云
何剑营
邱雷
乐涵波
LU Chang-chang;HUANG Yan-yan;XU Li-yun;HE Jian-ying;QIU lei;LE Han-bo(Cell and Molecular Biology Laboratory,Zhoushan Hospital,Zhejiang Zhoushan 316021,China;Hematology,Zhoushan Hospital,Zhejiang Zhoushan 316021,China;Cardio-Thoracic Surgery,Zhoushan Hospital,Zhejiang Zhoushan 316021,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2019年第2期192-200,共9页
Acta Anatomica Sinica
基金
浙江省科技厅项目(LGF18H160008
2015C37024
2017C37047)
舟山市科技局项目(2014C31065
2016C13037)
舟山市卫生局项目(2017G02)
