目的 探讨脂多糖(LPS)对人肺腺癌上皮A549细胞和人子宫颈癌上皮Hela细胞短的上腭、肺及鼻咽上皮克隆1(short palate, lung and nasal epithelium clone 1, SPLUNC1)蛋白表达的影响。方法 将两类细胞各随机分为对照组、实验组,CCK-8法观察LPS与细胞增殖活性的时间-反应和剂量-反应关系,筛选各自的最佳效应浓度和作用时间,以LPS最佳效应浓度和作用时间处理后,免疫组化和免疫荧光定量法检测两类细胞SPLUNC1蛋白相对表达量的变化。结果 LPS对A549细胞的最佳效应浓度为20 mg/L,最佳效应时间为12 h,对Hela细胞的最佳效应浓度为40 mg/L,12~24 h为其最佳效应时间窗。SPLUNC1在两类细胞中均呈阳性表达,LPS处理后表达均显著上调(P＜0.05),两类细胞间比较,LPS处理后Hela细胞SPLUNC1相对表达量高于A549细胞(P＜0.05),较对照的增量值(Δ)也明显大于A549细胞(P＜0.05)。结论 SPLUNC1在宫颈癌细胞中存在表达,可能是肺腺癌和宫颈癌病变中的保护性因子,具有潜在的临床应用价值。
Abstract
Objective To explore the influence of LPS on SPLUNC1 in Hela cells and A549 cells.Methods The two cell lines were cultured and divided into the control group and experimental group randomly. CCK-8 was used to study the time-response and dose-response relationship between LPS and proliferation and to screen the optimal concentration and action time of LPS in each cell line. After treatment with the optimal concentration and action time of LPS, the relative expression of SPLUNC1 was measured by immunohistochemical staining and immunofluorescence assay.Results The optimal concentration of LPS for A549 cells was 20 mg/L, and the optimal action time was 12 h. For Hela cells, the values were 40 mg/l and 12-24 h respectively. Both A549 cells and Hela cells expressed SPLUNC1. After LPS exposure,the expression of SPLUNC1 was up-regulated significantly(P＜0.05).Comparison between the two cell lines showed that after treatment with LPS the relative expression of SPLUNC1 in Hela cells was significantly higher than in A549 cells (P＜0.05), and the increment(Δ) of SPLUNC1 compared to the control in Hela cells was also significantly greater than in A549 cells(P＜0.05).Conclusions SPLUNC1 is expressed in cervical cancer. It can be a protective cell agent against lung adenocarcinoma and cervical cancer with potential clinical value.
关键词
短的上腭、肺及鼻咽上皮克隆1 /
A549细胞 /
Hela细胞 /
肿瘤 /
发病机制
Key words
SPLUNC1(short palate, lung and nasal epithelium clone 1) /
A549 cells /
Hela cells /
neoplasm /
pathogenesis
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Chen P, Guo X, Zhou H, et al. SPLUNC1 regulates cell progression and apoptosis through the miR-141-PTEN/p27 pathway, but is Hindered by LMP1[J]. PLoS One, 2013, 8 (3): e56929.
[2] 张晓丽,高宝德,刘海燕,等.重组SPLUNC1抑菌及调节NE活性的研究[J].中国预防兽医学报,2016,38(10): 823-826.
[3] Sayeed S, Nistico L, St Croix C, et al. Multifunctional role of human SPLUNC1 in pseudomonas aeruginosa infection [J]. Infect Immun, 2013, 81(1):285-291.
[4] Liu Y, Bartlett J A , Di M E, et al. SPLUNC1/BPIFA1 contributes to pulmonary host defense against Klebsiella pneumoniae respiratory infection[J]. Am J Pathol, 2013, 182(5):1519-1531.
[5] Teeguarden J G, Webb-Robertson B J, Waters K M, et al. Comparative proteomics and pulmonary toxicity of instilled singlewalled carbon nanotubes, crocidolite asbestos, and ultrafine carbon black in mice[J]. Toxicol Sci, 2011, 120(1): 123-135.
[6] Liu H, Zhang X,Wu J, et al. New insights on the palate, lung, and nasal epithelium clone (PLUNC) proteins: based on molecular and functional analysis of its homolog of YH1/SPLUNC1[J]. Exp Mol Pathol, 2016, 100(3):363-369.
[7] Chen W, Zheng R, Baade P D, et al. Cancer statistics in China, 2015[J]. CA Cancer J Clin, 2016, 66 (2):115-132.
[8] Crafton S M, Salani R. Beyond chemotherapy: an overview and review of targeted therapy in cervical cancer [J]. Clin Ther, 2016, 38(3):449-458.
[9] Pesic M, Greten F R. Inflammation and cancer: tissue regeneration gone awry[J]. Curr Opin Cell Biol, 2016, 43:55-61.
[10] Fernandes J V, Cobucci R N,Jatoba C A , et al. The role of the mediators of Iinflammation in cancer development[J]. Pathol Oncol Res, 2015, 21(3):527-534.
[11] Solinas G, Marchesi F, Garlanda C, et al. Inflammation-mediated promotion of invasion and metastasis [J]. Cancer Metastasis Rev, 2010, 29(2):243-248.
[12] Aggarwal B B, Sung B, Gupta S C. Advances in experimental medicine and biology[M]// Comes M , Teixeira AL, Coelho A, et al. The role of inflammation in lung cancer. Berlin: Springer Basel, 2014:1-23.
[13] 王婷婷, 周 瑾,黄百灵,等. HPV感染与宫颈癌预防的研究进展[J].现代生物医学进展,2014,14(20): 3988-3990.
[14] ACOG. Practice Bulletin No.157:cervical cancer screening and prevention[J]. Obstet Gynecol, 2016, 127(1):e1-e20.
[15] 王 爽,李文路,吕丽春,等.SPLUNC1基因在人体组织中的表达分布特征[J].南方医科大学学报,2016, 36(5): 617-621.
[16] Zhang L, Deng T, Li X, et al. MicroRNA-141 is involved in a nasopharyngeal carcinoma-related genes network[J]. Carcinogenesis, 2010, 3l(4):559-566.
[17] Benlloch S, Galbis-Caravajal J M, Alenda C, et al. Expression of molecular markers in mediastinal nodes from resected stageI non-small-cell lung cancer(NSCLC): prognostic impact and potential role as markers of occult micrometastases[J]. Ann Oncol, 2009, 20(1):91-97.
[18] Britto C J, Liu Q, Curran D R, et al. Short palate, lung, and nasal epithelial clone-1 is a tightly regulated airway sensor in innate and adaptive immunity[J]. Am J Respir Cell Mol Biol, 2013, 48(6): 717-724.
[19] Gakhar L, Bartlett J A, Penterman J, et al. PLUNC is a novel airway surfactant protein with anti-biofilm activity[J]. PLoS One, 2010, 5(2):e9098.
[20] Wei Y, Xia W, Ye X, et al. The antimicrobial protein short palate, lung, and nasal epithelium clone 1 (SPLUNC1) is differentially modulated in eosinophilic and noneosinophilic chronic rhinosinusitis with nasal polyps[J]. J Allergy Clin Immunol, 2014, 133(2):420-428.
[21] Musa M, Wilson K, Sun L, et al. Differential localisation of BPIFA1 (SPLUNC1) and BPIFB1 (SPLUNC1) in the nasal and oral cavities of mice[J]. Cell Tissue Res, 2012, 350 (3):455-464.
[22] 肖丁良,钟礼立,高喜容.脂多糖诱导SPLUNC1基因的表达[J].实用检验医师杂志,2017, 9(3): 170-172.
基金
山东省自然科学基金资助课题 (ZR2012CM008)
PDF(943 KB)
