摘要：
A549 cells were treated with different concentrations of anlotinib to create anlotinib‐resistant A549 cells (A549/anlotinib cells). miR‐181a‐3p mimics were transfected into A549/anlotinib cells. Meanwhile, A549 and A549/anlotinib cells were treated with β‐sitosterol at different concentrations. Cell Counting Kit‐8 (CCK‐8) was used to measure cell proliferation. The apoptosis level was detected by flow cytometry. Real‐time fluorescence quantitative PCR was used to detect the expression of miR‐181a‐3p. miR‐181a‐3p interaction with H/ACA ribonucleoprotein assembly factor (SHQ1) was predicted by miRDB and TargetScan Human databases and verified by luciferase reporter assay. The expressions of SHQ1, activating transcription factor 6 (ATF6) and glucose regulated protein 78 (GRP78) were detected by western blot. Our results show that β‐Sitosterol markedly promoted anlotinib‐resistant A549 cells apoptosis and inhibited the cell proliferation via activating SHQ1/UPR signaling via inhibiting miR‐181a‐3p. Abstract Anlotinib is used for the treatment of advanced non‐small cell lung cancer; however, the emergence of drug resistance limits its clinical application. β‐sitosterol may also be used to treat lung cancer, but there have been no studies evaluating β‐sitosterol against anlotinib‐resistant lung cancer. The purpose of this study was to determine the mechanism by which β‐sitosterol enhances the sensitivity of lung cancer cells to anlotinib. A549 cells were treated with different concentrations of anlotinib to generate anlotinib‐resistant cells (A549/anlotinib cells). miR‐181a‐3p mimics were transfected into A549/anlotinib cells. A549 and A549/anlotinib cells were treated with β‐sitosterol at various concentrations. The Cell Counting Kit‐8 (CCK‐8) assay was used to measure cell proliferation. Apoptosis was assessed by flow cytometry. Real‐time quantitative PCR was used to measure the expression of miR‐181a‐3p. The interaction of miR‐181a‐3p with the H/ACA ribonucleoprotein assembly factor (SHQ1) was predicted using the miRDB and TargetScan Human databases and verified with a luciferase reporter assay. The expression of SHQ1, activating transcription factor 6 (ATF6), and glucose‐regulated protein 78 (GRP78) were measured by western blot analysis. β‐Sitosterol effectively suppressed A549/anlotinib cell proliferation and promoted apoptosis. SHQ1 is a downstream target of miR‐181a‐3p. The expression of miR‐181a‐3p was inhibited; however, SHQ1 expression was increased by β‐sitosterol treatment of A549/anlotinib cells. The inhibition of SHQ1, ATF6, and GRP78 protein expression by β‐sitosterol in A549/anlotinib cells was rescued by increased miR‐181a‐3p. β‐Sitosterol markedly promotes anlotinib‐resistant A549 cell apoptosis and inhibits cell proliferation by activating SHQ1/UPR signaling through miR‐181a‐3p inhibition.
作者机构：
[Yi, Jian; Wang, Feiying; Ding, Rongzhen; Dai, Aiguo] Department of Respiratory Diseases, Medical School, Hunan University of Chinese Medicine, Changsha, China;[Yi, Jian; Wang, Feiying; Ding, Rongzhen; Dai, Aiguo] Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine, Changsha, China;[Ding, Rongzhen; Dai, Aiguo] Department of Respiratory Medicine, First Affiliated Hospital, Hunan University of Chinese Medicine, Changsha, China;[Sang, Shuliu] Shanghai University of Traditional Chinese Medicine, Shanghai, China;[Yi, Jian] Hunan Academy of Chinese Medicine, Changsha, China
摘要：
BACKGROUND: Lung adenocarcinoma (LUAD) with Pulmonary arterial hypertension (PAH) shows a poor prognosis. Detecting related genes is imperative for prognosis prediction. METHODS: The gene expression profiles of LUAD and PAH were acquired from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) database, respectively. The co-expression modules associated with LUAD and PAH were evaluated using the Weighted Gene Co-Expression Network Analysis (WGCNA). The relationship between key gene expression with immune-cell infiltration and the tumor immune microenvironment (TIME) was evaluated. We confirmed the mRNA and protein levels in vivo and vitro. G6PD knockdown was used to conduct the colony formation assay, transwell invasion assay, and scratch wound assay of A549 cells. EDU staining and CCK8 assay were performed on G6PD knockdown HPASMCs. We identified therapeutic drug molecules and performed molecular docking between the key gene and small drug molecules. RESULTS: Three major modules and 52 overlapped genes were recognized in LUAD and PAH. We identified the key gene G6PD, which was significantly upregulated in LUAD and PAH. In addition, we discovered a significant difference in infiltration for most immune cells between high- and low-G6PD expression groups. The mRNA and protein expressions of G6PD were significantly upregulated in LUAD and PAH. G6PD knockdown decreased proliferation, cloning, and migration of A549 cells and cell proliferation in HPASMCs. We screened five potential drug molecules against G6PD and targeted glutaraldehyde by molecular docking. CONCLUSIONS: This study reveals that G6PD is an immune-related biomarker and a possible therapeutic target for LUAD and PAH patients.
摘要：
Objective Based on metabolomics,it explored the differential metabolites screened by Chinese herbal medicines(compound/single decoction pieces)for the treatment of coronary heart disease with blood stasis syndrome in different species and summarized the biomarkers and related metabolic pathways after the intervention of Chinese herbal medicines.Methods The metabonomics study of traditional Chinese medicine(compound/single decoction pieces)in the treatment of coronary heart dis-ease with blood stasis syndrome was retrieved from CNKI,Wanfang database,VIP database,EMBASE,PubMed,the Cochrane Li-brary and web of science database.The differential metabolites included in the study were summarized.The metabolites were anno-tated by HMDB,PubChem subs and KEGG,and the metabolite path visualization was analyzed by metPA network software.Re-sults A total of eight publications were included in this study,and a total of 89 differential metabolites were counted after counting the recurring metabolites in both species that were mainly involved in 132 metabolic pathways.Through the analysis of pathway to-pology and the statistics of repeated metabolic pathways,16 metabolic pathways with P values less than 0.05 were selected as the metabolic pathways of traditional Chinese medicine(compound/single decoction pieces)in the treatment of coronary heart disease with blood stasis syndrome in different species.For the metabolites enriched in various differential metabolic pathways,the metab-olites repeatedly screened in different studies included phenylalanine,glutamine,glycine,citric acid,choline,creatine,lactic acid,β-glucose,α-glucose and arachidonic acid.Conclusion The herbs(compound/single decoction pieces)used for the treatment of coronary heart disease with blood stasis syndrome in various studies were represented by Xuefu Zhuyu Decoction(血府逐瘀汤),Taohong Siwu Decoction(桃红四物汤),Yangxin Tongmai Formula(养心通脉方)and Suhexiang(Styrax).Bioinformatics analysis based on metabolomics of Chinese herbal medicines(compound/single decoction pieces)for the treatment of coronary heart disease with blood stasis showed that the mechanism of action involves various aspects of glucose metabolism,amino acid metabolism,energy metabolism,choline metabolism and fatty acid metabolism,in which the basic tricarboxylic acid cycle is in-volved.The differential metabolites related to coronary heart disease with blood stasis syndrome,including lactic acid,glucose,cit-ric acid,arachidonic acid,creatine,phenylalanine,choline,glutamine and glycine,were summarized.
摘要：
OBJECTIVES: Statistics on the rate of unconventional lymph node metastases (ULNM) at the time of one-stage radical surgery in tongue cancer patients. To assess whether an extended neck dissection group with additional removal of ULNs has a lower rate of neck recurrence compared to the traditional neck dissection group. MATERIALS AND METHODS: A total of 336 patients with TSCC who underwent radical surgery were recruited and underwent traditional or extended neck dissection. Compared to traditional neck dissection, the aim of extended neck dissection is designed to additional resect ULNs. RESULTS: In total, 180 patients underwent extended neck dissection, while 156 underwent traditional neck dissection. The incidence of ULNM was 11.67% (21/180) in patients treated with extended neck dissection. The incidence of ipsilateral neck recurrence was 9.49% and 0.56% in patients who underwent traditional and extended neck dissection, respectively (p = 0.0001). CONCLUSIONS: Extended neck dissection is effective for preventing neck recurrence in TSCC patients with ULNs. CLINICAL RELEVANCE: ULNM may be the main cause of neck recurrence after neck dissection in patients with tongue cancer. A better prognosis may be achieved by additional resection of ULNs on the basis of traditional neck dissection.
期刊：
Journal of stomatology, oral and maxillofacial surgery,2024年:101836
通讯作者：
Zhu, Keke
作者机构：
[Zhu, Zibing; Tan, Jin; Zheng, Tao; Tan, Yisi] Department of Stomatology, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China;[Zheng, Tao] Hunan University of Chinese Medicine, Changsha, Hunan, China;[Zhu, Xuan; Liu, Chengyong] Hunan University of Chinese Medicine, Changsha, Hunan, China;[Zhou, Rong] Changsha Hospital for Maternal and Child Health Care, Changsha, Hunan, China;[Zhu, Keke] Department of Stomatology, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China. Electronic address: Zhukeke_HNUCM@163.com
通讯机构：
[Zhu, Keke] D;Department of Stomatology, The First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China. Electronic address:
关键词：
Cytokines;Head and neck neoplasms;Thyroid diseases
摘要：
INTRODUCTION: The established association between thyroid disorders (TD) and its two main subtypes-hyperthyroidism and hypothyroidism-and the incidence of oral and oropharyngeal cancer (OCPC) has been substantiated. However, the direct causal relationship and potential intermediary mechanisms linking these conditions have not been clearly defined in prior studies. MATERIAL & METHODS: This study employed univariate Mendelian randomization (MR) analysis to explore those relationship. Instrumental variables from genome-wide association study (GWAS) datasets for TD (n = 218,792), hyperthyroidism (n = 460,499), hypothyroidism (n = 213,990), and OCPC (n = 12,619), along with 41 intermediary inflammatory cytokines (n = 8293), were analyzed. Inverse variance weighting (IVW) method assessed the causal relationships, while summary MR analysis with pQTL datasets from decode and 91 inflammatory cytokines explored the cytokines' roles as biomarkers and therapeutic targets for OCPC. Multivariable MR (MVMR) analysis quantified the mediation effect of these cytokines in the TD-OCPC relationship. RESULTS: UVMR analysis provided strong evidence for a causal relationship between TD (OR = 1.376, 95 % CI = 1.142-1.656, p = 0.001), hyperthyroidism (OR = 1.319, 95 % CI=1.129-1.541, p = 0.001), hypothyroidism (OR = 1.224, 95 % CI = 1.071-1.400, p = 0.003), and the risk of OCPC. CXCL9 was identified as a significant intermediary in mediating the risk of OCPC from TD and its two subtypes (OR = 1.218, 95 % CI = 1.016-1.461, P = 0.033), suggesting its potential as a predictive biomarker for OCPC. MVMR analysis further revealed that CXCL9 mediated 7.94 %, 14.4 %, and 18 % of the effects of TD, hyperthyroidism, and hypothyroidism on OCPC risk, respectively. DISCUSSION: This study not only elucidated the potential causal relationships between TD including its two subtypes and OCPC risk, but also highlighted CXCL9 as a pivotal mediator in this association.
