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脂多糖和脂磷壁酸通过骨桥蛋白/整合素β3调节PI3K/AKT信号通路，以促进非小细胞肺癌的恶性进展。

Lipopolysaccharide and lipoteichoic acid regulate the PI3K/AKT pathway through osteopontin/integrin β3 to promote malignant progression of non-small cell lung cancer.

作者信息

Zhang Miao, Sun Yi, Zhang Yan, Wang Zhen, Wang Zhao-Yi, Ming Xi-Yue, Guo Zhen-Dong

机构信息

Department of Oncology, Shijiazhuang People's Hospital, Shijiazhuang, China.

Beijing Goldstandard Medicine Independent Clinical Laboratory Co. Ltd., Beijing, China.

出版信息

J Thorac Dis. 2023 Jan 31;15(1):168-185. doi: 10.21037/jtd-22-1825. Epub 2023 Jan 16.

DOI:10.21037/jtd-22-1825
PMID:36794132
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9922606/
Abstract

BACKGROUND: Lung cancer (LC) is a malignancy with one of the highest mortality rates. Respiratory microbiota is considered to play a key role in the development of LC, but the molecular mechanisms are rarely studied. METHODS: We used lipopolysaccharide (LPS) and lipoteichoic acid (LTA) to study human lung cancer cell lines PC9 and H1299. The gene expression of CXC chemokine ligand (CXCL)1/6, interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The Cell-Counting Kit 8 (CCK-8) was used to analyze cell proliferation. Transwell assays were performed to analyze cell migration ability. Flow cytometry was used to observe cell apoptosis. Western blot and qRT-PCR were used to analyze the expression of secreted phosphoprotein 1 (), toll-like receptor (TLR)-2/4, and NLR family pyrin domain containing 3 (NLRP3) to determine the mechanism of LPS + LTA. We evaluated the effect of LPS + LTA on cisplatin sensibility by analyzing cell proliferation, apoptosis, and caspase-3/9 expression levels. We observed the proliferation activity, apoptosis, and migration ability of cells in which had been transfected small interfering (si) negative control (NC) and integrin β3 siRNA. Then the mRNA expression level and protein expression of PI3K, AKT, and ERK were analyzed. Finally, the nude mouse tumor transplantation model was conducted to verify. RESULTS: We studied that in two cell lines, the expression level of inflammatory factors in LPS+LTA group was significantly higher than that in single treatment group (P<0.001). We explored LPS + LTA combined treatment group significantly increased the expression of NLRP3 and genes and proteins. LPS + LTA + Cisplatin group could significantly reduce the inhibitory effect of LPS on cell proliferation (P<0.001), reduce the apoptosis rate (P<0.001) and significantly reduce the expression levels of caspase-3/9 (P<0.001) compared with Cisplatin group. Finally, we verified that LPS and LTA could increase osteopontin (OPN)/integrin β3 expression and activate the PI3K/AKT pathway to promote malignant progression of LC studies. CONCLUSIONS: This study provides a theoretical basis for further exploration of the influence of lung microbiota on NSCLC and the optimization of LC treatment in the future.

摘要

背景：肺癌（LC）是死亡率最高的恶性肿瘤之一。呼吸道微生物群被认为在肺癌的发生发展中起关键作用，但分子机制鲜有研究。 方法：我们使用脂多糖（LPS）和脂磷壁酸（LTA）研究人肺癌细胞系PC9和H1299。通过定量实时聚合酶链反应（qRT-PCR）分析CXC趋化因子配体（CXCL）1/6、白细胞介素（IL）-6、IL-8和肿瘤坏死因子（TNF）-α的基因表达。使用细胞计数试剂盒8（CCK-8）分析细胞增殖。进行Transwell实验分析细胞迁移能力。采用流式细胞术观察细胞凋亡。使用蛋白质免疫印迹法和qRT-PCR分析分泌型磷蛋白1（）、Toll样受体（TLR）-2/4和含NLR家族pyrin结构域3（NLRP3）的表达，以确定LPS + LTA的作用机制。通过分析细胞增殖、凋亡和半胱天冬酶-3/9表达水平，评估LPS + LTA对顺铂敏感性的影响。我们观察了转染小干扰（si）阴性对照（NC）和整合素β3 siRNA的细胞的增殖活性、凋亡和迁移能力。然后分析PI3K、AKT和ERK的mRNA表达水平和蛋白质表达。最后，进行裸鼠肿瘤移植模型验证。 结果：我们研究发现，在两种细胞系中，LPS+LTA组炎症因子表达水平显著高于单一处理组（P<0.001）。我们探究LPS + LTA联合处理组显著增加了NLRP3及相关基因和蛋白的表达。与顺铂组相比，LPS + LTA +顺铂组可显著降低LPS对细胞增殖的抑制作用（P<0.001），降低凋亡率（P<0.001），并显著降低半胱天冬酶-3/9的表达水平（P<0.001）。最后，我们验证LPS和LTA可增加骨桥蛋白（OPN）/整合素β3表达并激活PI3K/AKT途径，促进肺癌的恶性进展 研究。 结论：本研究为进一步探索肺微生物群对非小细胞肺癌的影响及未来肺癌治疗的优化提供了理论依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/617b143065ed/jtd-15-01-168-f11.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/9be0805c1780/jtd-15-01-168-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/997149182445/jtd-15-01-168-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/08339e30d5db/jtd-15-01-168-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/4b9872cdd5e0/jtd-15-01-168-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/74887f283133/jtd-15-01-168-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/d2955f105009/jtd-15-01-168-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/aaa0f1def2dd/jtd-15-01-168-f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/3ea63351768b/jtd-15-01-168-f9.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/193cddb1e18a/jtd-15-01-168-f10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7efc/9922606/617b143065ed/jtd-15-01-168-f11.jpg

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[1]
Age-specific microbiota in altering host inflammatory and metabolic signaling and metabolome based on sex.

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[2]
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