m iR30a体外干预乳腺癌腋窝肿大淋巴结中肿瘤干细胞相关基因表达和侵袭力的变化

目的：探讨肿瘤干细胞在乳腺癌患者腋窝肿大淋巴结中微转移灶及微小RNA30a（miR30a）对其侵袭能力的影响,初步探讨miRNAs抗乳腺癌治疗的可行性。方法从乳腺癌患者腋窝肿大淋巴结中分离、培养肿瘤干细胞样乳腺癌细胞。合成miR30a寡核苷酸片段,应用腺病毒将该片段转染人原代乳腺癌细胞,同时设乳腺癌MDA‐MB‐231细胞株为试验对照,每组细胞均设空载体组、空白对照组,以荧光显微镜评估转染效率。以 T ransw ell小室体外侵袭试验检测转染前后肿瘤细胞增殖和侵袭力的变化。以Western blot分别检测ALDH1、Vimentin和N‐Cadherin蛋白表达。结果 Transwell小室体外侵袭试验显示空白对照组中原代乳腺癌细胞较 MDA‐MB‐231细胞株侵袭力强,其侵袭指数分别为（75．3±3．2）％,（58．4±2．8）％,两者差异有统计学意义（P＜0．05）,而转染miR30a后这两种细胞体外侵袭力均明显减弱,其侵袭指数分别为（21．4±1．9）％,（28．2±2．3）％,与各自空白对照组相比差异有统计学意义（P＜0．05）。结论乳腺癌患者部分腋窝增生肿大的淋巴结中ALDH表达增高,可能存在肿瘤微转移,术中应完全清扫为宜。miR30a抑制了肿瘤干性基因表达和侵袭能力。     

益气活血法通过miR216b/Beclin1介导萎缩性胃炎癌前病变自噬的机制

目的：通过观察益气活血法对萎缩性胃炎癌前病变(PLGC)模型小鼠微RNA216b(miR216b)/自噬关键分子酵母Atg6同系物1(Beclin1)信号通路的影响,探讨其干预PLGC的自噬作用机制。方法：将75只健康雄性SPF级昆明种小鼠随机分为空白组(12只)及造模组(63只),造模组小鼠每日采用1-甲基-3-硝基-1-亚硝基胍(MNNG)溶液(150 mg·L^-1)自由饮用及灌胃,同时每日灌胃雷尼替丁溶液(0.03 g·kg^-1),造模12周。按照随机对照表将造模小鼠分为模型组、益气组(黄芪3.5 g·kg^-1)、活血组(三七粉0.7 g·kg^-1)、益气活血组(黄芪3.5 g·kg^-1+三七粉0.7 g·kg^-1)及叶酸组(2 mg·kg^-1)。予相应药物灌胃8周后取材,苏木素-伊红(HE)染色观察小鼠胃黏膜病理改变,蛋白免疫印迹法(Western blot)检测胃组织微管相关蛋白1轻链3(LC3)Ⅰ、LC3Ⅱ、Beclin1...     

MiR‐483‐5p and miR‐139‐5p promote aggressiveness by targeting N‐myc downstream‐regulated gene family members in adrenocortical cancer

Adrenocortical carcinoma (ACC) is a tumor with poor prognosis in which overexpression of a panel of microRNAs has been associated with malignancy but a very limited number of investigations on their role in ACC pathogenesis have been conducted. We examined the involvement of miR‐483‐5p and miR‐139‐5p in adrenocortical cancer aggressiveness. Using bioinformatics predictions and mRNA/miRNA expression profiles, we performed an integrated analysis to identify inversely correlated miRNA–mRNA pairs in ACC. We identified N‐myc downstream‐regulated gene family members 2 and 4 (NDRG2 and NDRG4) as targets of miR‐483‐5p and miR‐139‐5p, respectively. NDRG2 and NDRG4 expressions were inversely correlated respectively with miR‐483‐5p and miR‐139‐5p levels in aggressive ACC samples from two independent cohorts of 20 and 44 ACC. Moreover, upregulation of miR‐139‐5p and downregulation of NDRG4 demonstrated a striking prognostic value. A direct interaction between miR‐483‐5p or miR‐139‐5p and their targets was demonstrated in reporter assays. Downregulation of miR‐483‐5p or miR‐139‐5p in the ACC cell lines NCI‐H295R and SW13 increased NDRG2 or NDRG4 mRNA and protein expression, compromised adrenocortical cancer cell invasiveness and anchorage‐independent growth. MiR‐483‐5p or miR‐139‐5p overexpression and NDRG2 or NDRG4 inhibition produce similar changes, which are rescued by NDRG2 or NDRG4 ectopic expression. We established that key factors mediating epithelial‐to‐mesenchymal transition are downstream effectors of miR‐483‐5p/NDRG2 and miR‐139‐5p/NDRG4 pathways. Collectively, our data show for the first time that miR‐483‐5p/NDRG2 and miR‐139‐5p/NDRG4 axes promote ACC aggressiveness, with potential implications for prognosis and therapeutic interventions in adrenocortical malignancies.     

Increased T‐helper 17 cell differentiation mediated by exosome‐mediated microRNA‐451 redistribution in gastric cancer infiltrated T cells

MicroRNA (miR)‐451 is a cell metabolism‐related miRNA that can mediate cell energy‐consuming models by several targets. As miR‐451 can promote mechanistic target of rapamycin (mTOR) activity, and increased mTOR activity is related to increased differentiation of T‐helper 17 (Th17) cells, we sought to investigate whether miR‐451 can redistribute from cancer cells to infiltrated T cells and enhance the distribution of Th17 cells through mTOR. Real‐time PCR was used for detecting expression of miR‐451 in gastric cancer, tumor infiltrated T cells and exosomes, and distribution of Th17 was evaluated by both flow cytometry and immunohistochemistry (IHC). Immunofluorescence staining was used in monitoring the exosome‐enveloped miR‐451 from cancer cells to T cells with different treatments, and signaling pathway change was analyzed by western blot. miR‐451 decreased significantly in gastric cancer (GC) tissues but increased in infiltrated T cells and exosomes; tumor miR‐451 was negatively related to infiltrated T cells and exosome miR‐451. Exosome miR‐451 can not only serve as an indicator for poor prognosis of post‐operation GC patients but is also related to increased Th17 distribution in gastric cancer. miR‐451 can redistribute from cancer cells to T cells with low glucose treatment. Decreased 5′ AMP‐activated protein kinase (AMPK) and increased mTOR activity was investigated in miR‐451 redistributed T cells and the Th17 polarized differentiation of these T cells were also increased. Exosome miR‐451 derived from tumor tissues can serve as an indicator for poor prognosis and redistribution of miR‐451 from cancer cells to infiltrated T cells in low glucose treatment can enhance Th17 differentiation by enhancing mTOR activity.