Down-Regulated miR-30a in Clear Cell Renal Cell Carcinoma Correlated with Tumor Hematogenous Metastasis by Targeting Angiogenesis-Specific DLL4

Background

Endothelial DLL4 plays an important role in controlling of tumor angiogenesis, which is required for tumor invasive growth and metastasis. However, the regulation of DLL4 in clear cell renal cell carcinoma (ccRCC) has not yet been systematically elucidated.

Methodology

We performed bioinformatical analysis to explore miRNAs targeting DLL4. miR-30a was selected as a representative to validate its functional association in endothelial cell. Then, the expressions of DLL4 and mature miR-30a from 90 cases of ccRCC and 28 cases of nonmatched adjacent non-tumor tissues were measured by quantitative real-time PCR. Finally, the expression of miR-30a was correlated with DLL4 expression, tumor features (metastatic condition and microvessel density), and patient metastasis-free survival. The univariate and multivariate analyses were performed to select the risk factors associated with hematogenous metastasis, respectively.

Principal Findings

miR-30a negatively regulated DLL4 and inhibited the proliferation and migration of endothelial cells. DLL4 was up-regulated in ccRCC and further increased in hematogenous metastatic cases, while miR-30a was down-regulated in tumor tissues and further decreased in hematogenous metastatic ccRCC (student t test, all p<0.05). Additionally, expression of miR-30a was inversely correlated with expression of DLL4 and microvessel density (linear correlation analysis, both p<0.05). Low-level miR-30a also indicated a higher probability of developing metastasis (log-rank test, p = 0.010). Most importantly, miR-30a expression was an independent predictor of ccRCC hematogenous metastasis by the univariate analysis and binary logistic regression model (both p<0.05).

Conclusions

Down-regulated miR-30a in ccRCC was associated with tumor hematogenous metastasis through increasing microvessel density by targeting angiogenesis-specific DLL4.     

Impact of miR-21, miR-126 and miR-221 as Prognostic Factors of Clear Cell Renal Cell Carcinoma with Tumor Thrombus of the Inferior Vena Cava

Clear cell renal cell carcinoma (ccRCC) characterized by a tumor thrombus (TT) extending into the inferior vena cava (IVC) generally indicates poor prognosis. Nevertheless, the risk for tumor recurrence after nephrectomy and thrombectomy varies. An applicable and accurate prediction system to select ccRCC patients with TT of the IVC (ccRCC/TT) at high risk after nephrectomy is urgently needed, but has not been established up to now. To our knowledge, a possible role of microRNAs (miRs) for the development of ccRCC/TT or their impact as prognostic markers in ccRCC/TT has not been explored yet. Therefore, we analyzed the expression of the previously described onco-miRs miR-200c, miR-210, miR-126, miR-221, let-7b, miR-21, miR-143 and miR-141 in a study collective of 74 ccRCC patients. Using the expression profiles of these eight miRs we developed classification systems that accurately differentiate ccRCC from non-cancerous renal tissue and ccRCC/TT from tumors without TT. In the subgroup of 37 ccRCC/TT cases we found that miR-21, miR-126, and miR-221 predicted cancer related death (CRD) accurately and independently from other clinico-pathological features. Furthermore, a combined risk score based on the expression of miR-21, miR-126 and miR-221 was developed and showed high sensitivity and specificity to predict cancer specific survival (CSS) in ccRCC/TT. Using the combined risk score we were able to classify ccRCC/TT patients correctly into high and low risk cases. The risk stratification by the combined risk score (CRS) will benefit from further cohort validation and might have potential for clinical application as a molecular prediction system to identify high- risk ccRCC/TT patients.     

Exosomal miR-146a-5p and miR-155-5p promote CXCL12/CXCR7-induced metastasis of colorectal cancer by crosstalk with cancer-associated fibroblasts

C-X-C motif chemokine receptor 7 (CXCR7) is a newly discovered atypical chemokine receptor that binds to C-X-C motif chemokine ligand 12 (CXCL12) with higher affinity than CXCR4 and is associated with the metastasis of colorectal cancer (CRC). Cancer-associated fibroblasts (CAFs) have been known to promote tumor progression. However, whether CAFs are involved in CXCR7-mediated metastasis of CRC remains elusive. We found a significant positive correlation between CXCR7 expression and CAF activation markers in colonic tissues from clinical specimens and in villin-CXCR7 transgenic mice. RNA sequencing revealed a coordinated increase in the levels of miR-146a-5p and miR-155-5p in CXCR7-overexpressing CRC cells and their exosomes. Importantly, these CRC cell-derived miR-146a-5p and miR-155-5p could be uptaken by CAFs via exosomes and promote the activation of CAFs through JAK2–STAT3/NF-κB signaling by targeting suppressor of cytokine signaling 1 (SOCS1) and zinc finger and BTB domain containing 2 (ZBTB2). Reciprocally, activated CAFs further potently enhanced the invasive capacity of CRC cells. Mechanistically, CAFs transfected with miR-146a-5p and miR-155-5p exhibited a robust increase in the levels of inflammatory cytokines interleukin-6, tumor necrosis factor-α, transforming growth factor-β, and CXCL12, which trigger the epithelial–mesenchymal transition and pro-metastatic switch of CRC cells. More importantly, the activation of CAFs by miR-146a-5p and miR-155-5p facilitated tumor formation and lung metastasis of CRC in vivo using tumor xenograft models. Our work provides novel insights into CXCR7-mediated CRC metastasis from tumor–stroma interaction and serum exosomal miR-146a-5p and miR-155-5p could serve as potential biomarkers and therapeutic targets for inhibiting CRC metastasis.Subject terms: Cancer microenvironment, Colon cancer     

Down-Regulation of miR-129-5p Inhibits Growth and Induces Apoptosis in Laryngeal Squamous Cell Carcinoma by Targeting APC

miRNAs regulate gene expression and are key mediators of tumourigenesis. miR-129 has diverse effects in tumours, but its role in laryngeal squamous cell carcinoma (LSCC) remains unknown. This article focuses on the role of miR-129-5p in LSCC. We show miR-129-5p is upregulated in primary LSCC tumours and correlated with advanced disease. Down-regulating miR-129-5p suppressed cell proliferation and migration, and caused cell cycle arrest in Hep-2 cell lines. Downregulation of miR-129-5p alone is sufficient to induce apoptosis both in vivo and in vitro. Moreover, the growth of LSCC xenograft exposed to miR-129-5p antisense oligonucleotides (ASO) in BALB/c mice was markedly inhibited. In addition, we found that miR-129-5p targeted adenomatous polyposis coli (APC) to release inhibition of Wnt signalling causing cell growth and tumourigenesis. Our results suggest miR-129-5p functions as an oncogene in LSCC by repressing APC and is a potential therapeutic target for LSCC.     

Genetic Variants in Caveolin-1 and RhoA/ROCK1 Are Associated with Clear Cell Renal Cell Carcinoma Risk in a Chinese Population

Background

The RhoA/ROCK pathway and Caveolin-1 (Cav-1) participate in the process of tumorigenesis in numerous types of cancer. Up-regulation of RhoA/ROCK and Cav-1 expression is considered to be associated with the development and progression of clear cell renal cell carcinoma (ccRCC). We investigated the association between genetic variations of RhoA/ROCK and Cav-1 and the risk of ccRCC in the Chinese population.

Methods

Between May 2004 and March 2014, a total of 1,248 clear cell renal cell carcinoma cases and 1,440 cancer-free controls were enrolled in this hospital-based case-control study. Nine SNPs in RhoA/ROCK and Cav-1 were genotyped using the TaqMan assay.

Result

We found two SNPs (Cav-1 rs1049334 and ROCK1 rs35996865) were significantly associated with the increasing risk of ccRCC (P = 0.002 and P < 0.001 respectively). The analysis of combined risk alleles revealed that patients with 2–4 risk alleles showed a more remarkable growth of ccRCC risk than the patients with 0–1 risk alleles(OR = 1.66, 95%CI = 1.31–2.11, P < 0.001). Younger subjects (P = 0.001, OR = 1.83, 95%CI = 1.30–2.57), higher weight subjects (P = 0.001, OR = 1.76, 95%CI = 1.25–2.47), female subjects (P = 0.007, OR = 1.75, 95% CI = 1.17–2.62), nonsmokers (P < 0.001, OR = 1.67, 95%CI = 1.26–2.23), drinkers (P = 0.025, OR = 1.75, 95% CI = 1.07–2.85), subjects with hypertension (P = 0.025, OR = 1.75, 95% CI = 1.07–2.85) and diabetes (P = 0.026, OR = 4.31, 95% CI = 1.19–15.62) showed a stronger association between the combined risk alleles and the risk of ccRCC by using the stratification analysis. Furthermore, we observed higher Cav-1 mRNA levels in the presence of the rs1049334 A allele in normal renal tissues.

Conclusion

Our results indicate that the two SNPs (Cav-1 rs1049334 and ROCK1 rs35996865) and genotypes with a combination of 2–4 risk alleles were associated with the risk of ccRCC. The functional SNP rs1049334 may affect the risk of ccRCC by altering the expression of Cav-1 and the relevance between the risk effects and the functional impact of this polymorphism needs further validation.     

miR-150-5p Inhibits Hepatoma Cell Migration and Invasion by Targeting MMP14

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related mortality worldwide. Despite progress in diagnostics and treatment of HCC, its prognosis remains poor because the molecular mechanisms underlying hepatocarcinogenesis are not well understood. In the study, we focused on identifying the role of miRNAs in HCC progression. miRNA microarray was used to analyze the differentially expressed miRNAs, and the results were validated by qPCR. We found that the miR-150-5p expression is down-regulated in HCC tissues compared with pair non-tumor tissues. miR-150-5p expression is also decreased in metastatic cancer tissues compared with pair primary tissues, indicating that miR-150-5p may be involved in HCC metastasis. Functionally, miR-150-5p inhibition significantly promotes hepatoma cell migration and invasion, whereas miR-150-5p overexpression suppresses cancer cell migration and invasion in vitro. The matrix metalloproteinase 14 (MMP14) is identified as a new target gene of miR-150-5p. miR-150-5p markedly inhibits MMP14 expression in hepatoma cells, and miR-150-5p expression is negative correlation with MMP14 expression in vivo. More important, re-expression of MMP14 in hepatoma cells partially reverses the effect of miR-150-5p in inhibiting cell invasion.     

喉鳞状细胞癌组织miR-1207-5p、miR-186-5p表达水平与PI3K/Akt信号通路、临床病理特征和预后的关系

摘要 目的：探讨喉鳞状细胞癌（LSCC）组织miR-1207-5p、miR-186-5p表达水平与磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路、临床病理特征和预后的关系。方法：选取2017年1月至2020年1月南充市中心医院收治的120例LSCC患者,取手术切除的LSCC组织和癌旁组织。检测miR-1207-5p、miR-186-5p、PI3K mRNA、Akt mRNA表达。患者出院后随访3年,统计总生存（OS）和无复发生存（RFS）情况。分析miR-1207-5p、miR-186-5p与PI3K、Akt的相关性以及影响LSCC患者预后的因素。结果：LSCC组织miR-1207-5p、miR-186-5p表达低于癌旁组织（P＜0.05）,PI3K mRNA、Akt mRNA表达高于癌旁组织（P＜0.05）。LSCC组织miR-1207-5p、miR-186-5p表达与PI3K mRNA、Akt mRNA表达呈负相关（P＜0.05）。肿瘤直径≥1 cm、低分化、TNM分期Ⅲ期、颈部淋巴结转移LSCC组织中miR-1207-5p、miR-186-5p表达低于肿瘤直径＜1 cm、中高分化、TNM分期Ⅰ～Ⅱ期、无颈部淋巴结转移（P＜0.05）。miR-1207-5p低表达、miR-186-5p低表达LSCC患者3年总生存（OS）率和无复发生存（RFS）率低于miR-1207-5p高表达、miR-186-5p高表达LSCC患者（P＜0.05）。多因素COX回归分析显示TNM III期、颈部淋巴结转移是LSCC患者复发和死亡的危险因素（P＜0.05）,高miR-1207-5p、高miR-186-5p是保护因素（P＜0.05）。结论：LSCC组织中miR-1207-5p和miR-186-5p表达均下调,与LSCC恶性病理特征、PI3K/Akt信号通路激活以及低生存率有关。     

Expression of miR-142-5p in Peripheral Blood Mononuclear Cells from Renal Transplant Patients with Chronic Antibody-Mediated Rejection

In renal transplantation, the unresponsiveness of patients undergoing chronic antibody mediated rejection (CAMR) to classical treatment stress on the need for accurate biomarkers to improve its diagnosis. We aim to determine whether microRNA expression patterns may be associated with a diagnosis of CAMR. We performed expression profiling of miRNAs in peripheral blood mononuclear cells (PBMC) of kidney transplant recipients with CAMR or stable graft function. Among 257 expressed miRNAs, 10 miRNAs associated with CAMR were selected. Among them, miR-142-5p was increased in PBMC and biopsies of patients with CAMR as well as in a rodent model of CAMR. The lack of modulation of miR-142-5p in PBMC of patients with renal failure, suggests that its over-expression in CAMR was associated with immunological disorders rather than renal dysfunction. A ROC curve analysis performed on independent samples showed that miR-142-5p is a potential biomarker of CAMR allowing a very good discrimination of the patients with CAMR (AUC = 0.74; p = 0.0056). Moreover, its expression was decreased in PHA-activated blood cells and was not modulated in PBMC from patients with acute rejection, excluding a non-specific T cell activation expression. The absence of modulation of this miRNA in immunosuppressed patients suggests that its expression was not influenced by treatment. Finally, the analysis of miR-142-5p predicted targets under-expressed in CAMR PBMC in a published microarray dataset revealed an enrichment of immune-related genes. Altogether, these data suggest that miR-142-5p could be used as a biomarker in CAMR and these finding may improve our understanding of chronic rejection mechanisms.     

慢性牙周炎患者血清miR-205-5p、miR-28-5p的表达与诊断价值研究

摘要 目的：探讨慢性牙周炎（CP）患者血清微小核糖核酸（miRNA）-205-5p、miR-28-5p表达与牙周指标、炎症反应的关系及诊断价值。方法：选取2021年1月～2022年12月我院口腔科收治的102例CP患者为CP组,根据病情严重程度分为轻度组31例、中度组37例、重度组34例,另选取同期我院65名体检健康者为对照组。采用实时荧光定量聚合酶链式反应（PCR）检测血清miR-205-5p、miR-28-5p表达,酶联免疫吸附法检测白细胞介素（IL）-1β、IL-6、肿瘤坏死因子-α（TNF-α）水平,检查各组牙周指标[菌斑指数（PLI）、牙龈出血指数（BI）、附着丧失（AL）、探诊深度（PD）]。CP患者血清miR-205-5p、miR-28-5p表达与牙周指标和炎症指标的相关性采用Pearson相关性分析。受试者工作特征（ROC）曲线分析血清miR-205-5p、miR-28-5p表达诊断CP的价值。结果：与对照组比较,CP组血清miR-205-5p、miR-28-5p表达降低,PLI、BI、AL、PD和IL-1β、IL-6、TNF-α水平升高（P＜0.05）。轻度组、中度组、重度组血清miR-205-5p、miR-28-5p表达依次降低,PLI、BI、AL、PD和IL-1β、IL-6、TNF-α水平依次升高（P＜0.05）。Pearson相关性分析显示,CP患者血清miR-205-5p、miR-28-5p表达与PLI、BI、AL、PD、IL-1β、IL-6、TNF-α呈负相关（P＜0.05）。ROC曲线分析显示,血清miR-205-5p联合miR-28-5p诊断CP的曲线下面积分别为0.885大于miR-205-5p、miR-28-5p单独检测的0.792、0.790。结论：CP患者血清miR-205-5p、miR-28-5p低表达,与牙周指标、炎症反应密切相关,血清miR-205-5p、miR-28-5p的表达情况可能成为CP辅助诊断指标,且miR-205-5p联合miR-28-5p诊断CP的价值较高。     

肝细胞癌组织miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路和预后的关系研究

摘要 目的：探讨肝细胞癌组织中微小核糖核酸（miR）-124-3p、miR-212-5p表达与磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路和预后的关系。方法：选择2017年9月至2019年9月徐州医科大学附属医院肝胆外科收治的93例肝细胞癌患者,采用实时荧光定量聚合酶链反应（qRT-PCR）检测肝细胞癌组织中miR-124-3p、miR-212-5p、PI3K 信使RNA（mRNA）、Akt mRNA的表达。Pearson相关性分析miR-124-3p、miR-212-5p表达与PI3K/Akt信号通路相关mRNA表达的相关性。绘制Kaplan-Meier生存曲线,Log-Rank检验不同miR-124-3p、miR-212-5p表达肝细胞癌患者3年总生存率（OS）的差异。结果：肝细胞癌组织中miR-124-3p表达低于癌旁组织（P＜0.05）,miR-212-5p、PI3K mRNA、Akt mRNA表达高于癌旁组织（P＜0.05）。肝细胞癌组织中miR-124-3p表达与PI3K mRNA、Akt mRNA表达呈负相关（P＜0.05）,miR-212-5p表达与PI3K mRNA、Akt mRNA表达呈正相关（P＜0.05）。Ⅱa期、低分化患者肝细胞癌组织中miR-124-3p表达低于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）,miR-212-5p表达高于Ⅰa-Ⅰb期、中高分化患者（P＜0.05）。随访期间死亡37例, miR-124-3p低表达组3年OS为42.55%,低于miR-124-3p高表达组的78.26%（P＜0.05）,miR-212-5p高表达组3年OS为50.00%,低于miR-212-5p低表达组的71.11%（P＜0.05）。结论：肝细胞癌组织中miR-124-3p表达下调,miR-212-5p表达上调,且与肝细胞癌PI3K/Akt信号通路激活,PI3K mRNA和Akt mRNA高表达,低分化,CNLC分期Ⅱa期以及低OS有关。     

Circulating miR-23b-3p,miR-145-5p and miR-200b-3p are potential biomarkers to monitor acute pain associated with laminitis in horses

Circulating microRNAs (miRNAs) are emerging as promising biomarkers for several disorders and related pain. In equine practice, acute laminitis is a common disease characterised by intense pain that severely compromises horse welfare. Recently, the Horse Grimace Scale (HGS), a facial expression-based pain coding system, was shown to be a valid welfare indicator to identify pain linked to acute laminitis. The present study aimed to: determine whether miRNAs can be used as biomarkers for acute pain in horses (Equus caballus) affected by laminitis; integrate miRNAs to their target genes and to categorise target genes for biological processes; gather additional evidence on concurrent validity of HGS by investigating how it correlates to miRNAs. Nine horses presenting acute laminitis with no prior treatment were recruited. As control group, nine healthy horses were further included in the experimental design. Samples were collected from horses with laminitis at admission before any treatment (‘pre-treatment’) and 7 days after routine laminitis treatment (‘post-treatment’). The expression levels of nine circulating miRNAs, namely hsa-miR-532-3p, hsa-miR-219-5p, mmu-miR-134-5p, mmu-miR-124a-3p, hsa-miR-200b-3p, hsa-miR-146a-5p, hsa-miR-23b-3p, hsa-miR-145-5p and hsa-miR-181a-5p, were detected and assessed as potential biomarkers of pain by quantitative PCR using TaqMan® probes. The area under the receiver operating curve (AUC) was then used to evaluate the diagnostic performance of miRNAs. Molecular data were integrated with HGS scores assessed by one trained treatment and time point blind veterinarian. The comparative analysis demonstrated that the levels of miR-23b-3p (P=0.029), miR-145-5p (P=0.015) and miR-200b-3p (P=0.023) were significantly higher in pre-treatment and the AUCs were 0.854, 0.859 and 0.841, respectively. MiR-200b-3p decreased after routine laminitis treatment (P=0.043). Combining two miRNAs in a panel, namely miR-145-5p and miR-200b-3p, increased efficiency in distinguishing animals with acute pain from controls. In addition, deregulated miRNAs were positively correlated to HGS scores. Computational target prediction and functional enrichment identified common biological pathways between different miRNAs. In particular, the glutamatergic pathway was affected by all three miRNAs, suggesting a crucial role in the pathogenesis of pain. In conclusion, the dynamic expression of circulating miR-23b-3p, miR-145-5p and miR-200b-3p was detected in horses with acute laminitis and miRNAs can be considered potentially promising pain biomarkers. Further studies are needed in order to assess their relevancy in other painful conditions severely compromising horse welfare. An important implication would be the possibility to use them for the concurrent validation of non-invasive indicators of pain in horses.     

miR-369-3p靶向FGF9信号通路调控肝癌细胞增殖与侵袭

目的:阐明脂代谢相关mi R-369-3p在肝癌(hepatocellular carcinoma, HCC)病理组织中的表达特征及其与临床预后的相关性,解析mi R-369-3p在HCC细胞中的作用性质及相应分子机理。方法:实时定量PCR法检测mi R-369-3p在HCC病理组织和细胞中的表达变化,并通过Spearman's法分析mi R-369-3p表达水平与临床病理资料相关性;CCK-8检测、Transwell穿透小室实验和裸鼠荷瘤实验检测敲低内源性mi R-369-3p表达后对HCC细胞增殖和侵袭性的影响作用;利用生物信息学分析、瞬时转染、定点突变和荧光素酶报告基因活性检测分析mi R-369-3p对纤维母细胞生长因子9(fibroblast growth factor 9,FGF9)的转录后调控作用。结果:mi R-369-3p在HCC病理组织(n=68)和细胞中异常低表达,且此低表达趋势与HCC患者预后呈显著负相关关系(x~2=6.907,P=0.0086);敲低内源性mi R-369-3p可显著促进HCC细胞的增殖、侵袭性;参与调控这一抑瘤效应的关键分子机制可能是miR-369-3p与FGF9的3'-UTR区直接结合,在转录后水平抑制FGF9 m RNA的稳定性,进而抑制后者表达水平。结论:miR-369-3p可通过靶向FGF9信号在转录后调控水平负性调控肝癌细胞增殖和侵袭过程,在肝癌发生和进展过程中发挥关键抑癌作用。     

miR-1915 and miR-1225-5p Regulate the Expression of CD133, PAX2 and TLR2 in Adult Renal Progenitor Cells

Adult renal progenitor cells (ARPCs) were recently identified in the cortex of the renal parenchyma and it was demonstrated that they were positive for PAX2, CD133, CD24 and exhibited multipotent differentiation ability. Recent studies on stem cells indicated that microRNAs (miRNAs), a class of noncoding small RNAs that participate in the regulation of gene expression, may play a key role in stem cell self-renewal and differentiation. Distinct sets of miRNAs are specifically expressed in pluripotent stem cells but not in adult tissues, suggesting a role for miRNAs in stem cell self-renewal. We compared miRNA expression profiles of ARPCs with that of mesenchymal stem cells (MSCs) and renal proximal tubular cells (RPTECs) finding distinct sets of miRNAs that were specifically expressed in ARPCs. In particular, miR-1915 and miR-1225-5p regulated the expression of important markers of renal progenitors, such as CD133 and PAX2, and important genes involved in the repair mechanisms of ARPCs, such as TLR2. We demonstrated that the expression of both the renal stem cell markers CD133 and PAX2 depends on lower miR-1915 levels and that the increase of miR-1915 levels improved capacity of ARPCs to differentiate into adipocyte-like and epithelial-like cells. Finally, we found that the low levels of miR-1225-5p were responsible for high TLR2 expression in ARPCs. Therefore, together, miR-1915 and miR-1225-5p seem to regulate important traits of renal progenitors: the stemness and the repair capacity.     

miR-305-3p和miR-71-5p通过调控谷胱甘肽代谢途径参与斜纹夜蛾应对植物次生物质

昆虫和植物在长期进化过程中形成相互作用的关系。其中植物次生物质是植物防御昆虫的主要机制,而昆虫则以解毒酶来应对。为了发现可用于害虫防治的miRNA,通过对农业害虫斜纹夜蛾Spodoptera litura进食芥菜Brassica juncea后中肠的测序分析,获得了一系列差异表达的miRNA。通过生物信息学分析,发现斜纹夜蛾miR-305-3p靶向了谷胱甘肽代谢中的谷氨酸半胱氨酸连接酶的催化亚基(Slgclc),这是一个谷胱甘肽从头合成的限速酶;而miR-71-5p靶向调控gclc和解毒酶表达的转录因子SlNrf2。用芥菜的次生物质吲哚-3-甲醇处理斜纹夜蛾Spli-221细胞株后,实时荧光定量PCR证明Slgclc和SlNrf2表达上调,miR-305-3p和miR-71-5p表达下调。分别在斜纹夜蛾Spli-221细胞中超表达miR-305-3p及miR-71-5p mimics,Slgclc或SlNrf2转录水平下调,并且miR-71-5p mimic处理抑制了SlNrf2下游基因Slgclc和解毒酶谷胱甘肽S-转移酶(GSTs)的表达。结果表明:miR-305-3p和miR-71-5p响应植物次生物质而分别促进Slgclc和SlNrf2表达。然而双荧光素酶实验显示miR-305-3p并不与Slgclc直接结合,miR-71-5p也不与SlNrf2直接结合,推测miR-305-3p和miR-71-5p可能间接调控Slgclc和SlNrf2的表达。研究结果表明,斜纹夜蛾miR-305-3p和miR-71-5p通过调控解毒酶GSTs表达及其底物谷胱甘肽的生成,而参与昆虫抵抗植物次生物质。