丝氨酸蛋白酶抑制因子Kazal 1型在人肺腺癌组织表达上调并促进肺腺癌细胞的增殖

doi:10.16098/j.issn.0529-1356.2019.02.009

摘要/Abstract

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。

关键词: 肺腺癌, 丝氨酸蛋白酶抑制因子Kazal 1型, 细胞增殖, 实时定量聚合酶连反应, 人

Abstract:

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 enzyme-linked 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.

Key words: Lung adenocarcinoma, Serine protease inhibitor Kazal type 1, Cell proliferation, Real-time PCR, Human

陆畅畅黄燕燕续力云何剑营邱雷乐涵波. 丝氨酸蛋白酶抑制因子Kazal 1型在人肺腺癌组织表达上调并促进肺腺癌细胞的增殖[J]. 解剖学报, 2019, 50(2): 192-200.

LU Chang-chang HUANG Yan-yan XU Li-yun HE Jian-ying QIU lei LE Han-bo. Up-regulation of serine protease inhibitor Kazal type 1 expression in human lung adenocarcinoma and promotion of proliferation of cancer cells [J]. Acta Anatomica Sinica, 2019, 50(2): 192-200.

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