针对肺腺癌癌细胞中的 KRAS、p38α 和 NF-κB：RNA 干扰与化学抑制剂联合使用的效果。
Targeting the KRAS, p38α, and NF-κB in lung adenocarcinoma cancer cells: The effect of combining RNA interferences with a chemical inhibitor.
机构信息
Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Science, Tabriz, Iran.
Students Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
出版信息
J Cell Biochem. 2019 Jun;120(6):10670-10677. doi: 10.1002/jcb.28357. Epub 2019 Jan 17.
BACKGROUND
Lung cancer is the leading cause of cancer-related death with less than 5-year survival rate for both men and women worldwide. KRAS (Kirsten rat sarcoma), nuclear factor-κB (NF-κB), and mitogen-activated protein kinase (MAPK) signaling pathways have a critical role in the proliferation and progression of various cancers, including lung cancer. The p38 MAPK plays a different role in various tissue hence show a tissue-dependent behavior. It acts as an oncogene in some tissues while plays as a tumor suppressor in some other tissues. Also, KRAS and NF-κB act as an oncogene in various cancer. This study was dedicated to analyzing the combined effect of NF-κB inhibitor, specific KRAS, and p38α small interfering RNA (siRNA) in A549 cell line.
MATERIALS AND METHODS
The cytotoxic effects of p38α siRNA, KRAS siRNA, and NF-κB inhibitor were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) assay. Relative p38α, KRAS, and NF-κB messenger RNA (mRNA) levels were measured by quantitative reverse-transcription polymerase chain reaction. Induction of apoptosis by treatments was measured by fluorescence-activated cell sorting (FACS) analysis.
RESULTS
The expression of mRNA related to p38α and KRAS genes was reduced to 23.4% and 26.7%, respectively, after treatment with specific siRNAs. Also, MTT assay showed that the cell viability after treatment with p38α siRNA, KRAS siRNA, NF-κB inhibitor and their combination was reduced. FACS results indicated that p38α siRNA, KRAS siRNA, and NF-κB inhibitor, and their combination, reduced the population of live cells in comparison with the population of untreated control cells (99.5%). The results are expressed as mean ± SD (n = 3); *P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 vs control group.
CONCLUSION
The results of this study indicated that p38α, KRAS, and NF-κB signaling pathways might play an important role in the development and growth of lung cancer and might be a potential therapeutic target for treatment of lung cancer.
背景
肺癌是导致癌症相关死亡的主要原因，全球男性和女性的 5 年生存率均较低。KRAS（Kirsten 大鼠肉瘤）、核因子-κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）信号通路在各种癌症（包括肺癌）的增殖和进展中起着关键作用。p38MAPK 在不同的组织中发挥不同的作用，表现出组织依赖性。在某些组织中作为癌基因，而在某些其他组织中作为肿瘤抑制因子。此外，KRAS 和 NF-κB 在各种癌症中作为癌基因。本研究旨在分析 NF-κB 抑制剂、特异性 KRAS 和 p38α 小干扰 RNA（siRNA）在 A549 细胞系中的联合作用。
材料和方法
使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法测定 p38αsiRNA、KRASsiRNA 和 NF-κB 抑制剂的细胞毒性作用。通过定量逆转录聚合酶链反应（qRT-PCR）测量相对 p38α、KRAS 和 NF-κB 信使 RNA（mRNA）水平。通过荧光激活细胞分选（FACS）分析测量处理诱导的细胞凋亡。
结果
用特异性 siRNA 处理后，p38α 和 KRAS 基因相关 mRNA 的表达分别降低至 23.4%和 26.7%。此外，MTT 测定表明，用 p38α siRNA、KRAS siRNA、NF-κB 抑制剂及其组合处理后，细胞活力降低。FACS 结果表明，与未经处理的对照细胞群相比，p38α siRNA、KRAS siRNA、NF-κB 抑制剂及其组合减少了活细胞群体（99.5%）。结果表示为平均值±标准差（n=3）；P<0.05；* P<0.01；*** P<0.001；**** P<0.0001 与对照组相比。
结论
本研究结果表明，p38α、KRAS 和 NF-κB 信号通路可能在肺癌的发生和生长中起重要作用，可能是治疗肺癌的潜在治疗靶点。
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