Circulating miRNA-126, -145 and -155 levels in Mexican women exposed to inorganic arsenic via drinking water

The aim of this research was to investigate circulating expression levels of three miRNAs (miR-126, miR-155, and miR-145) proposed as predictive CVD biomarkers in Mexican women exposed to inorganic arsenic via drinking water. Mean UAs concentration of 19.5 ± 14.0 μg/g creatinine was found after urine samples were analyzed (n = 105). Significant associations between UAs levels and serum expression levels of miR-155 (p < 0.05) and miR-126 (p < 0.05) were observed after adjustment for assessed co-variables. Alterations in the serum expression levels of miR-155 and miR-126 may be associated with the onset and development of cardiovascular diseases, hence miRNAs could be proposed as prognostic CVD biomarkers. Data found in this study are of concern and risk reduction plans are necessary for the assessed communities to prevent cardiovascular events in this population of women.     

miR-155联合超声对分化型甲状腺癌淋巴结转移的诊断价值分析

目的　探讨微小RNA-155（miR-155）联合超声对分化型甲状腺癌淋巴结转移的诊断价值。方法　选取接受治疗的分化型甲状腺癌患者112例作为甲状腺癌组,另选取同期收治的结节性甲状腺肿患者30例作为良性对照组。采用实时荧光定量PCR（qPCR）检测2组患者血清及组织中miR-155的相对表达量,分析分化型甲状腺癌患者血清及组织中miR-155的表达与临床病理参数的关系。采用受试者工作特征（ROC）曲线分析术前超声以及血清miR-155对分化型甲状腺癌淋巴结转移的诊断价值。结果　甲状腺癌组的血清及组织中miR-155的相对表达量均明显高于良性对照组（P＜0.01）,有淋巴结转移、肿瘤直径＞2 cm、TNM分期Ⅲ+Ⅳ期的患者血清及组织中miR-155的相对表达量分别高于无淋巴结转移、肿瘤直径≤2 cm、TNM分期Ⅰ+Ⅱ期的患者（均P＜0.05）。ROC结果显示,超声以及血清miR-155对分化型甲状腺癌淋巴结转移均有一定的诊断价值,曲线下面积分别为：0.839（95%CI：0.760～0.919）、0.837（95%CI：0.763～0.912）,但漏诊率和误诊率均超过10%。两者联合应用后对分化型甲状腺癌淋巴结转移的诊断价值可得到进一步的提升,曲线下面积为0.912（95%CI：0.851～0.973）,漏诊率和误诊率也降至10%以下。结论　血清miR-155联合超声对分化型甲状腺癌淋巴结转移的诊断价值较高,具有一定的临床应用价值。     

Lead (Pb) exposure is associated with changes in the expression levels of circulating miRNAS (miR-155, miR-126) in Mexican women

The environmental contamination with lead (Pb) is considered a critical issue worldwide. Therefore, this study aimed to evaluate the expression levels of circulating miRNAs (miR-155, miR-126, and miR-145) in Mexican women exposed to Pb. Blood lead levels (BLL) were assessed in enrolled women (n = 190) using an atomic absorption method. Also, serum miRNAs expression levels were quantified through a real-time PCR assay. A mean BLL of 10.5 ± 4.50 μg/dL was detected. Overexpression of miR-155 was detected in highly exposed women. Besides, a significant simple positive relationship (p < 0.05) was found between BLL and serum miR-155 expression levels. Additionally, a significant inverse correlation (p < 0.05) was determined between BLL and serum miR-126 expression levels, as downregulation of miR-126 expression levels was observed in highly exposed women. The findings in this study are the concern, as epigenetic changes detected may represent a connection between health illnesses and Pb exposure.     

Decreased Expression of Plasma MicroRNA in Patients with Methamphetamine (MA) Use Disorder

Recent research have revealed that circulating miRNAs may offer noninvasive biomarkers for human disease, offering the prospect for earlier diagnosis, and improved precision of diagnoses. The diagnoses of drug use disorders is still mainly based on subjective report and no objective biomarkers available. Many animal and cell studies found that miRNAs were involved in substance use disorders, including alcohol, morphine, cocaine and amphetamine use disorders. However, no study on circulating miRNAs for drug use disorders so far. We investigated the differential expression of plasma miRNAs in 124 patients with methamphetamine (MA) use disorders. Based on the preliminary results from microarray screen, plasma expression of 6 candidate miRNAs were measured by Quantitative real-time RT-PCR. We found that the expression of miR181a, miR15b, miR- let-7e, miR- let-7d in plasma were decreased compared to normal controls. The expression of the altered miRNAs were negative correlated with drug use frequencies in past months. Our findings suggested that miR-181a, miR-15b, miR-let-7e and miR-let-7d may play a potential role in the pathology of MA use disorder, and could serve as a potential peripheral biomarker for MA use disorder when confirmed by future studies. Further study are needed to elucidate the molecular mechanism modulated by miRNAs and explore potential novel intervention targets.     

MicroRNA-mediated suppression of P-glycoprotein by quantum dots in lung cancer cells

The interactions between adenosine triphosphate-binding cassette (ABC) transporters and nano-sized materials are attracting increasing attention, due to their great potential in overcoming the multidrug resistance (MDR) phenomena in cancer treatment. However, the inner mechanisms involved in the interactions are largely unknown. In this study, two commercial quantum dots (QDs), CdSe/ZnS-MPA and CdSe/ZnS-GSH, were tested for their interactions with P-glycoprotein (P-gp), as well as the relating mechanisms in lung cancer (A549) cells. Both QDs significantly suppressed the gene and protein expressions of P-gp in A549 cells. To explain this, the gene expressions of nine relating microRNAs (miRNAs) were evaluated. The results indicated a shared up-regulation of miR-34b and miR-185 by both QDs. Furthermore, mimics and inhibitors of miR-34b and miR-185 significantly enhanced and suppressed the gene and protein expressions of P-gp, respectively, confirming the modulatory function of these two miRNAs on P-gp. Interestingly, expressions of both miRNAs were suppressed during treatment with Cd²⁺ and doxorubicin, which induced the expression of P-gp, indicating the universality of these miRNAs-related mechanisms. Thus, as miR-34b and miR-185 participated in the suppression of P-gp functions in A549 cells they could be interesting targets for the treatment of lung cancer.     

非吸烟女性肺腺癌组织中 miRNAs 的表达与表皮生长因子受体关系的研究

目的 探讨非吸烟女性肺腺癌(ADC)组织中 miR-155、miR-16、miR-25 及 miR-133a 的表达与表皮生长因子受体（EGFR）的关系及其临床意义。 方法 112 例非吸烟女性 ADC 患者按 EGFR 类型分为 EGFR 野生型组（n= 51）和 EGFR 突变型组（n=61）。 用实时荧光定量 PCR 定量检测 2 组 ADC 组织中 4 种 miRNAs 的表达量, 比较 2 组 4 种 miRNAs 表达水平的差异。 4 种 miRNAs 表达量以平均数分层为高表达组和低表达组, 比较 2 亚组患者间的生存差异。 按年龄（≤60 岁, ＞ 60 岁）、EGFR 类型和 miRNAs 的表达量分层, 进行 Kaplan-Meier 生存分析和 Cox 比例风险回归模型检验。 结果 EGFR 突变型组 miR-25 的表达水平高于 EGFR 野生型组（P < 0.05）。 不同年龄分层、不同 EGFR 类型组、miR-155、miR-16 及 miR-133a 表达量患者的生存率差异无统计学意义, miR-25 低表达者的生存率较高表达者更高（P < 0.05）。 在 EGFR 突变型组中 4 种 miRNAs 的表达量与 ADC 预后无关（P > 0.05）; 在 EGFR 野生型组中 miR-16、miR-25、miR-133a 低表达者较高表达者生存率更高（P < 0.05）。 Cox 比例风险回归分析显示, miR-25 高表达是非吸烟女性 ADC 患者预后死亡的独立危险因素（P < 0.05）。 结论 miR-25 是非吸烟女性 ADC 的独立预后指标。 miR-25 在非吸烟女性 ADC, 特别是在 EGFR 野生型的患者中表达升高, 可能提示患者预后不佳。     

血清外泌体miR-155、miR-222表达对乳腺癌术后复发转移的预测价值

目的 探究血清外泌体miR-155、miR-222表达对乳腺癌术后复发转移的预测价值。方法 选取2016年1月至2017年6月于南通市中医院收治的乳腺癌病人72例作为乳腺癌组,同期选取确诊为乳腺纤维瘤病人40例为对照组。根据乳腺癌病人术后是否发生复发转移分为复发转移组27例和未复发转移组45例。利用外泌体试剂盒提取血清中外泌体,采用荧光定量PCR技术检测外泌体miR-155、miR-222水平。采用受试者工作特征（ROC）曲线评价血清外泌体miR-155、miR-222水平对乳腺癌病人术后复发转移的诊断价值。结果 研究组血清外泌体miR-155(1.46±0.62)、miR-222(1.87±0.79)水平均明显高于对照组[（0.98±0.03）、（1.02±0.02）]（均P<0.05）;复发转移组病人血清外泌体miR-155(1.68±0.47)、miR-222(2.13±0.53)水平均明显高于未复发转移组[（1.33±0.24）、（1.71±0.26）]（均P<0.05）;临床病理分析结果显示,乳腺癌病人血清外泌体miR-155、miR-222表达与TNM分期、淋巴...     

Cigarette smoking substantially alters plasma microRNA profiles in healthy subjects

Circulating microRNAs (miRNAs) are receiving attention as potential biomarkers of various diseases, including cancers, chronic obstructive pulmonary disease, and cardiovascular disease. However, it is unknown whether the levels of circulating miRNAs in a healthy subject might vary with external factors in daily life. In this study, we investigated whether cigarette smoking, a habit that has spread throughout the world and is a risk factor for various diseases, affects plasma miRNA profiles. We determined the profiles of 11 smokers and 7 non-smokers by TaqMan MicroRNA array analysis. A larger number of miRNAs were detected in smokers than in non-smokers, and the plasma levels of two-thirds of the detected miRNAs (43 miRNAs) were significantly higher in smokers than in non-smokers. A principal component analysis of the plasma miRNA profiles clearly separated smokers and non-smokers. Twenty-four of the miRNAs were previously reported to be potential biomarkers of disease, suggesting the possibility that smoking status might interfere with the diagnosis of disease. Interestingly, we found that quitting smoking altered the plasma miRNA profiles to resemble those of non-smokers. These results suggested that the differences in the plasma miRNA profiles between smokers and non-smokers could be attributed to cigarette smoking. In addition, we found that an acute exposure of ex-smokers to cigarette smoke (smoking one cigarette) did not cause a dramatic change in the plasma miRNA profile. In conclusion, we found that repeated cigarette smoking substantially alters the plasma miRNA profile, interfering with the diagnosis of disease or signaling potential smoking-related diseases.     

Exposure to biomass smoke is associated with an increased expression of circulating miRNA-126 and miRNA-155 in Mexican women: a pilot study

Household air pollution has been associated as a risk factor for cardiovascular diseases (CVD). Therefore, the aim of this study was to assess the expression of vascular inflammation regulators miR-126 and miR-155 in plasma from women that cook with wood and women that cook with liquid petroleum gas (LPG). A cumulative index of exposure to smoke (CIES) was estimated, urinary 1-hydroxypyrene (1-OHP) levels were quantified and miRNAs expression levels were determined by quantitative real-time PCR (qRT-PCR). Biochemical clinical parameters were also evaluated. The average values for CIES and 1-OHP were 140?±?86.8?hours-years (12.0–270?hours-years) and 0.52?±?0.45?µmol/mol creatinine, respectively. miR-126 and miR-155 expression levels were significantly higher (p?<?0.01) in the wood users compared to LPG users. Besides, we found a significant association (p?<?0.01) between miR-126 and miR-155 expression levels and CIES and urinary 1-OHP concentrations. These results contribute to the current evidence about the cardiovascular risk related to biomass smoke exposure, from an epigenetic level.     

Circulating microRNA profiles altered in mice after 28 d exposure to perfluorooctanoic acid

Perfluorooctanoic acid (PFOA) is a stable man-made compound with many industrial and commercial uses. Recently, however, concern has been raised that it may induce various toxicological effects such as hepatotoxicity, immunotoxicity, and developmental toxicity. Because levels of circulating microRNAs (miRNAs) can be altered in several clinical diseases, they may serve as potential novel biomarkers. Here, we explored differences in the profiles of circulating miRNAs in mice after PFOA exposure. Using TaqMan miRNA arrays, we determined that the levels of 24 circulating miRNAs were altered in mice dosed with PFOA at 1.25 mg/kg/d and 73 were altered in mice dosed with 5 mg/kg/d. Eight miRNAs were further validated using TaqMan Real-Time PCR assays. Results were consistent with those obtained from the TaqMan miRNA arrays, except for miR-199a-3p. The most remarkable of the circulating miRNAs (miR-26b-5p and miR-199a-3p) were also up-regulated in the serum of occupational workers in our previous epidemiological study. We also found similar patterns in mice exposed to PFOS. These results demonstrated that circulating miRNA profiles were altered after exposure to high concentrations of PFOA and miR-28-5p, miR-32-5p, miR-122-5p, miR-192-5p, and miR-26b-5p in serum may be linked to effects of PFOA, especially in occupationally exposed people.     

Targeted Stage-Specific Inflammatory microRNA Profiling in Urine During Disease Progression in Experimental Autoimmune Encephalomyelitis: Markers of Disease Progression and Drug Response

Recently, microRNAs (miRNAs) have been implicated in regulating neuroinflammatory and demyelinative responses in multiple sclerosis (MS) and its mouse model of experimental autoimmune encephalomyelitis (EAE). miRNAs have also been studied as biomarkers of disease pathology and drug-response in MS. However, no complete miRNA profiling at various stages of EAE disease has been examined, especially in the urine. We carried out a systematic analysis of miRNAs in the urine exosomes as well as in the plasma and spinal cord at pre-onset, onset and peak stages of EAE established in the chronic B6 mice model. For the first time, we provide evidence that urine exosomes can be a specific and sensitive source of miRNA biomarkers for all 3 stages of EAE disease. In a significant observation, we observed that miR-155-5p expression increased in urine exosomes, plasma and spinal cord 6 days before the onset of disease, suggesting its early involvement in the pathology of EAE disease. We also analyzed the effect of Glatiramer acetate (GA; copaxone) treatment, an approved treatment for MS patients, in modulating miRNA expression at the peak of EAE disease. We identified miR-155-5p, miR-27a-3p, miR-9-5p and miR-350-5p as putative GA-treatment responsive miRNA biomarkers. Since, EAE is a mainly CD4 cells mediated disease, we also examined the above set of miRNAs and found to be significantly altered in T cells polarized to Th1 and Th17 phenotype, similar to urine exosomes. Thus, urine exosome miRNAs hold the potential to be defined as novel accessible stage-specific biomarkers of EAE (MS) disease as well as treatment response.     

miR-25对食管癌EC109细胞侵袭和迁移能力的 影响及临床意义

目的 探索 miR-25 对食管癌侵袭和迁移能力的影响以及作为食管癌诊断生物标志物的潜力。方法 （1）荧光定量PCR（qPCR）检测54例早期食管癌患者癌组织和癌旁组织中miR-25表达水平。（2）人食管癌细胞EC109 分为miR-25 mimic组、NC mimic组、miR-25 inhibitor组和NC inhibitor组。转染相应序列后,qPCR检测miR-25转染 效率。Transwell实验检测过表达或敲低miR-25对EC109细胞侵袭和迁移的影响。Targetscan数据库预测miR-25的 靶基因,选定靶基因盐诱导激酶1（SIK1）基因。Western blot 和双荧光素酶报告实验鉴定miR-25与SIK1的靶向关 系。（3）EC109 细胞分为 pcDNA 3.1 组、SIK1 过表达组和 miR-25+SIK1 过表达组。Transwell 实验检测 miR-25 靶向 SIK1对EC109细胞侵袭和迁移能力的影响。（4）提取食管癌患者（54例）和健康对照者（54例）的血浆外泌体,比较2 组血浆外泌体中miR-25的相对表达量,分析食管癌患者癌组织和血浆外泌体中miR-25的相关性。结果 （1）食管 癌组织中 miR-25 相对表达水平高于癌旁组织。（2）过表达 miR-25 后,EC109 细胞侵袭和迁移能力增强,而敲低 miR-25 表达后,细胞侵袭和迁移能力下降。Western blot 结果显示,过表达 miR-25 后,SIK1 蛋白表达下降;反之, 敲低miR-25后SIK1蛋白表达升高。（3）Transwell实验显示,与pcDNA 3.1组相比,SIK1过表达组EC109细胞侵袭和 迁移的细胞数量减少,而miR-25和SIK1联合作用后,EC109侵袭和迁移的细胞数量较SIK1过表达组增多。（4）食管 癌血浆外泌体样本中miR-25的表达量高于健康对照,且miR-25在食管癌组织中的表达与血浆外泌体中的相对表达 量呈正相关。结论 miR-25可能通过靶向调控SIK1来促进食管癌细胞的侵袭和迁移,且miR-25有作为食管癌早 期诊断生物标志物的潜力。     

Plasma microRNAs are sensitive indicators of inter-strain differences in the severity of liver injury induced in mice by a choline- and folate-deficient diet

MicroRNAs (miRNAs) are a class of small, conserved, tissue-specific regulatory non-coding RNAs that modulate a variety of biological processes and play a fundamental role in the pathogenesis of major human diseases, including nonalcoholic fatty liver disease (NAFLD). However, the association between inter-individual differences in susceptibility to NAFLD and altered miRNA expression is largely unknown. In view of this, the goals of the present study were (i) to determine whether or not individual differences in the extent of NAFLD-induced liver injury are associated with altered miRNA expression, and (ii) assess if circulating blood miRNAs may be used as potential biomarkers for the noninvasive evaluation of the severity of NAFLD. A panel of seven genetically diverse strains of inbred male mice (A/J, C57BL/6J, C3H/HeJ, 129S/SvImJ, CAST/EiJ, PWK/PhJ, and WSB/EiJ) were fed a choline- and folate-deficient (CFD) diet for 12 weeks. This diet induced liver injury in all mouse strains; however, the extent of NAFLD-associated pathomorphological changes in the livers was strain-specific, with A/J, C57BL/6J, and C3H/HeJ mice being the least sensitive and WSB/EiJ mice being the most sensitive. The morphological changes in the livers were accompanied by differences in the levels of hepatic and plasma miRNAs. The levels of circulating miR-34a, miR-122, miR-181a, miR-192, and miR-200b miRNAs were significantly correlated with a severity of NAFLD-specific liver pathomorphological features, with the strongest correlation occurring with miR-34a. These observations suggest that the plasma levels of miRNAs may be used as biomarkers for noninvasive monitoring the extent of NAFLD-associated liver injury and susceptibility to NAFLD.     

MicroRNA-34a-5p as a promising early circulating preclinical biomarker of doxorubicin-induced chronic cardiotoxicity

Cardiotoxicity is a serious adverse effect of an anticancer drug, doxorubicin (DOX), which can occur within a year or decades after completion of therapy. The present study was designed to address a knowledge gap concerning a lack of circulating biomarkers capable of predicting the risk of cardiotoxicity induced by DOX. Profiling of 2083 microRNAs (miRNAs) in mouse plasma revealed 81 differentially expressed miRNAs 1 week after 6, 9, 12, 18, or 24 mg/kg total cumulative DOX doses (early-onset model) or saline (SAL). Among these, the expression of seven miRNAs was altered prior to the onset of myocardial injury at 12 mg/kg and higher cumulative doses. The expression of only miR-34a-5p was significantly (false discovery rate [FDR] < 0.1) elevated at all total cumulative doses compared with concurrent SAL-treated controls and showed a statistically significant dose-related response. The trend in plasma miR-34a-5p expression levels during DOX exposures also correlated with a significant dose-related increase in cardiac expression of miR-34a-5p in these mice. Administration of a cardioprotective drug, dexrazoxane, to mice before DOX treatment, significantly mitigated miR-34a-5p expression in both plasma and heart in conjunction with attenuation of cardiac pathology. This association between plasma and heart may suggest miR-34a-5p as a potential early circulating marker of early-onset DOX cardiotoxicity. In addition, higher expression of miR-34a-5p (FDR < 0.1) in plasma and heart compared with SAL-treated controls 24 weeks after 24 mg/kg total cumulative DOX dose, when cardiac function was altered in our recently established delayed-onset cardiotoxicity model, indicated its potential as an early biomarker of delayed-onset cardiotoxicity.     

微小RNA-20a、微小RNA-92a在乳腺癌诊断及预后评估中的应用

目的 分析微小RNA(miR)-20a、miR-92a在乳腺癌病人血清中的表达情况,并探讨其对乳腺癌诊断及预后评估的价值.方法 选取2012年1月至2013年5月中国人民解放军中部战区总医院首次确诊为乳腺癌并经手术治疗的病人89例为观察组,以80例同期健康体检者为健康对照组,采用实时荧光定量逆转录聚合酶链式反应(qRT...     

血清miR-122和miR-155在丙戊酸钠诱导小鼠仔鼠肝损伤中的表达

目的 探讨血清miR-122和miR-155在丙戊酸钠（VPA）致小鼠仔鼠肝损伤过程中的动态表达及其与肝损 伤的关系。方法 4周龄昆明小鼠按随机数字表法分为对照组30只和实验组56只。实验组又分为7组,分别给予 VPA 375 mg·kg-1·d-1连续灌胃1、3、5、7、14、21和28 d;对照组又分为5组,分别于1、7、14、21和28 d 给予1 mL生理盐 水灌胃,留取血清及肝组织。HE染色观察肝组织病理改变;全自动生化分析仪测定血清丙氨酸转氨酶（ALT）和天冬 氨酸转氨酶（AST）;实时荧光定量聚合酶链式反应（qPCR）检测血清miR-122和miR-155表达水平。结果 肝组织 病理学结果显示,实验组肝细胞出现不同程度的空泡变性。3、5、7 d时肝细胞胞浆内可见少量炎性细胞,21 d时大 量肝细胞变性坏死,28 d时大量肝细胞再生。血清ALT 与AST均呈现下降-升高的趋势。实验组给药后miR-122 表达水平下调,7 d 时降至最低点（下降了 81%）,之后上调;而 miR-155 的表达趋势与 miR-122 相反,7 d 时达到最 高点,是对照组的2.88倍。miR-122与ALT和AST呈正相关（rs分别为0.965、0.900,均P＜0.05）,miR-155与AST呈 负相关（rs=-0.811,P＜0.05）。结论 miR-122和miR-155参与了VPA药物性肝损伤的发生,有望成为VPA药物性肝 损伤的早期预警指标。     

慢病毒介导的microRNA-155过表达对肝癌细胞HepG2增殖的影响

摘要： 目的 通过构建慢病毒 （LV） 介导的miRNA-155 （miR-155） 过表达载体， 探讨上调miR-155对肝癌细胞系 HepG2增殖的影响。方法 针对目标序列合成特异性miR-155过表达片段并克隆入慢病毒载体pGCSHL-GFP， 将重组miR-155-hRNA-LV和阴性对照空病毒载体转染HepG2细胞， 建立稳定过表达miR-155的细胞系。细胞设过表达组 （H组）、 阴性对照组 （negative control， NC组） 和正常组 （normal， N组）。实时荧光聚合酶链式反应 （qRT-PCR） 方法检测各组miR-155和PTEN的表达水平； Western blot检测各组PTEN、 CyclinD1、 CyclinA1+A2蛋白表达情况； 细胞增殖-毒性检测 （CCK-8） 细胞增殖情况。结果 H组miR-155表达量明显高于其NC组及N组， 靶基因PTEN的表达量低于NC组及N组 （均P＜0.05）。H组PTEN蛋白的表达量明显低于NC组和N组， 而CyclinD1、 CyclingA1+A2蛋白的表达量明显高于NC组和N组 （均P＜0.05）。CCK-8结果显示3组12 h时吸光度值开始出现差异， 24 h、 48 h、 72 h H 组吸光度值均明显大于NC组和N组 （P＜0.05）， 但后2组差异无统计学意义。结论 过表达miR-155可促进肝癌细胞系HepG2细胞增殖。     

Analyses of the possible anti-tumor effect of yokukansan

The Kampo medicine yokukansan (YKS) has a wide variety of properties such as anxiolytic, anti-inflammatory and analgesic effects, and is also thought to regulate tumor suppression. In this study, we investigated the anti-tumor effect of YKS. We used Lewis lung carcinoma (LLC)-bearing mice that were fed food pellets containing YKS and then performed a fecal microbiota analysis, a microarray analysis for microRNAs (miRNAs) and an in vitro anti-tumor assay. The fecal microbiota analysis revealed that treatment with YKS partly reversed changes in the microbiota composition due to LLC implantation. Furthermore, a miRNA array analysis using blood serum showed that treatment with YKS restored the levels of miR-133a-3p/133b-3p, miR-1a-3p and miR-342-3p following LLC implantation to normal levels. A TargetScan analysis revealed that the epidermal growth factor receptor 1 signaling pathway is one of the major target pathways for these miRNAs. Furthermore, treatment with YKS restored the levels of miR-200b-3p and miR-200c-3p, a recognized mediator of cancer progression and controller of emotion, in the hypothalamus of mice bearing LLC. An in vitro assay revealed that a mixture of pachymic acid, saikosaponins a and d and isoliquiritigenin, which are all contained in YKS, exerted direct and additive anti-tumor effects. The present findings constitute novel evidence that YKS may exert an anti-tumor effect by reversing changes in the fecal microbiota and miRNAs circulating in the blood serum and hypothalamus, and the compounds found in YKS could have direct and additive anti-tumor effects.