miR-613对高尔基体磷蛋白3调控乳腺癌细胞增殖、迁移和侵袭的研究

目的研究miR-613靶向调控高尔基体磷蛋白3(GOLPH3)基因对乳腺癌MCF-7细胞的增殖、迁移和侵袭的机制。方法用脂质体技术分别将miR-NC、miR-613 mimic、anti-miR-NC、anti-miR-613、si-NC、si-GOLPH3、miR-613 mimic与pcDNA、miR-613mimic与pcDNA-GOLPH3转染至MCF-7细胞。用实时荧光定量聚合酶链反应检测miR-613和GOLPH3 mRNA的表达,用噻唑蓝法、Transwell法检测MCF-7细胞的增殖、迁移和侵袭。结果 HBL-100组和MCF-7组的miR-613表达量分别为1.03±0.09和0.37±0.04,GOLPH3 mRNA表达量分别为1.07±0.11和3.25±0.26,GOLPH3蛋白表达量分别为0.34±0.05和0.75±0.08,MCF-7组的上述指标与HBL-100组比较,差异均有统计学意义(均P<0.05)。miR-NC组和miR-613组72 h时细胞增殖率分别为(0.97±0.09)%和(0.63±0.06)%,迁移细胞数分别为(86.79±8.1...     

姜黄素通过上调miR-34a抑制人神经母细胞瘤SH-SY5Y细胞的恶性生物学行为

目的 探讨姜黄素对人神经母细胞瘤SH-SY5Y细胞恶性生物行为的影响及其可能作用机制。方法 CCK8法检测姜黄素(0、6.25、12.5、25、50和100μmol·L^-1)作用下细胞活力并筛选出后续姜黄素处理浓度(0、6.25、12.5、25μmol·L^-1),流式细胞法、划痕实验、Transwell实验和Western blot分别检测细胞凋亡、迁移、侵袭及相关蛋白水平。使用qRT-PCR法检测SH-SY5Y细胞miR-34a表达水平,构架miR-34a低表达SH-SY5Y细胞系(miR-34a inhibitor组)及其阴性对照(NC组)、对照组(Control组),三组细胞系均给予姜黄素(25μmol·L^-1)处理分别记为黄素+miR-34a inhibitor组、姜黄素+NC组、姜黄素组,另设置空白对照组,采用上述相同方法检测各组细胞凋亡、迁移、侵袭及相关蛋白水平和p-p65、p-IκBα蛋白水平。结果 当姜黄素干预浓度达到12.5μmol·L^-1时,SH-SY5Y细胞存活率显著低于无姜黄...     

miR-96在人乳腺癌中表达及其对乳腺癌细胞侵袭和迁移活性的影响

目的研究miR-96在人乳腺上皮细胞株和乳腺病变组织中表达状况及其对人乳腺癌细胞增殖、侵袭和迁移的影响。方法抽提4株人乳腺上皮细胞系和28例人乳腺病变新鲜组织总miRNA,实时定量PCR方法检测它们miR-96的表达状况;脂质体介导的转染方法将miR-96抑制物转染人乳腺癌MCF-7细胞株;通过MTS试剂盒检测细胞增殖能力,Transwell侵袭和迁移实验检测细胞侵袭及迁移能力。结果miR-96在高侵袭乳腺癌细胞株中表达较低侵袭乳腺细胞明显下调(P<0.01);人乳腺癌组织中miR-96表达较乳腺良性病变组织明显下调(P<0.01);miR-96抑制物转染乳腺癌MCF-7后,乳腺癌细胞侵袭和迁移活力较阴性对照组细胞明显增强(P<0.05),而细胞增殖活力改变无统计学意义(P>0.05)。结论 miR-96在人乳腺癌细胞株和乳腺癌组织中存在表达下调,并对人乳腺癌细胞的侵袭及迁移能力可能存在一定负性调控作用。     

MAPKAPK5-AS1/miR-96对乳腺癌细胞转移的影响及机制

目的 探讨长链非编码RNA MAPKAPK5-AS1(LncRNA MAPKAPK5-AS1)/微小RNA-96(miR-96)对乳腺癌细胞增殖及迁移的影响及作用机制.方法 采用实时荧光定量聚合酶链反应(qRT-PCR)检测乳腺癌组织及癌旁组织中MAPKAPK5-AS1的表达;体外培养乳腺癌细胞MDA-MB-468,分...     

乳腺癌患者血清miR-135b、miR-194表达量与肿瘤恶性增殖及侵袭的相关性分析

目的 探讨乳腺癌患者血清中miR-135b与miR-194的表达量,并与肿瘤的恶性增殖与侵袭进行相关性分析.方法 选择2018年9月至2019年6月进行手术切除的乳腺癌患者40例,收集乳腺癌患者手术切除的乳腺癌组织、癌旁正常乳腺组织及患者进行手术之前的血清标本,同时选取同时期进行体检的健康者40例并收集血清.采用RT-...     

外源高表达miR-133 b抑制骨肉瘤的增殖与转移

目的 分析miR-133b对骨肉瘤细胞增殖与转移的影响.方法 qRT-PCR检测人骨肉瘤细胞系(MG-63)与人正常成骨细胞系(hFOB)中miR-133b的表达水平;运用MTT实验检测miR-133b对人骨肉瘤MG-63细胞的生长能力的作用;运用划痕实验检测miR-133b对人骨肉瘤MG-63细胞的迁移能力的作用;T...     

circ_0000264促进乳腺癌细胞增殖、迁移和侵袭

目的 探讨环状RNA circ_0000264在乳腺癌细胞增殖、迁移和侵袭中的功能及作用机制。方法 采用qRT-PCR检测乳腺癌组织和细胞系中circ_0000264的表达;利用si-circ_0000264构建敲低circ_000026的细胞模型;分别采用CCK-8、BrdU和Transwell实验检测细胞的增殖、迁移和侵袭;采用双荧光素酶报告基因实验验证circ_0000264与miR-622的靶向调控关系。结果 circ_0000264在乳腺癌组织和细胞中均呈高表达（P<0.05）。与对照组相比,敲低circ_0000264可抑制乳腺癌细胞的增殖、迁移和侵袭（P<0.05）。circ_0000264对miR-622具有吸附作用。下调miR-622可部分逆转敲低circ_0000264对乳腺癌细胞增殖和转移的抑制作用。结论 circ_0000264在乳腺癌进展过程中发挥促进作用;circ_0000264通过负向调控miR-622促进乳腺癌细胞的增殖、迁移和侵袭。     

Evodiamine inhibits malignant progression of ovarian cancer cells by regulating lncRNA-NEAT1/miR-152-3p/CDK19 axis

Evodiamine (EVO) has been demonstrated to promote apoptosis of ovarian cancer cells, and upregulate miR-152-3p level in colorectal cancer. Here, we explore part of the network mechanism of EVO and miR-152-3p in ovarian cancer. The bioinformatics website, dual luciferase reporter assay, and quantitative real-time polymerase chain reaction were applied to analyze the network among EVO, lncRNA, miR-152-3p, and mRNA. The effect and mechanism of EVO on ovarian cancer cells were determined using cell counting kit-8, flow cytometry, TUNEL, Western blot, and rescue experiments. As a result, EVO dose-dependently attenuated cell viability, induced G2/M phase arrest and apoptosis, promoted miR-152-3p level (4.5- or 2-fold changes), and inhibited expressions of NEAT1 (0.225- or 0.367-fold changes), CDK8 (0.625- or 0.571-fold changes), and CDK19 (0.25- or 0.147-fold changes) in OVCAR-3 and SKOV-3 cells. In addition, EVO decreased Bcl-2 expression, but increased the expressions of Bax and c-caspase-3. NEAT1 targeted miR-152-3p which bound to CDK19. The impacts of EVO on cell viability, cycle, apoptosis, and apoptosis-related proteins were partially reversed by miR-152-3p inhibitor, NEAT1 overexpression, or CDK19 overexpression. Furthermore, miR-152-3p mimic offset the effects of NEAT1 or CDK19 overexpression. The role of NEAT1 overexpression in the biological phenotype of ovarian cancer cells was counteracted by shCDK19. In conclusion, EVO attenuates ovarian cancer cell progression via the NEAT1-miR-152-3p-CDK19 axis.     

miR-187通过靶向胰岛素生长因子-1受体基因对人乳腺癌细胞增殖和凋亡的影响

目的 研究miR-187通过靶向胰岛素生长因子-1受体(IGF-1R)基因对人乳腺癌MDA-MB-231细胞增殖和凋亡的影响.方法 采用实时荧光定量PCR(qRT-PCR)检测人正常乳腺上皮细胞HBL-100和不同乳腺癌细胞中miR-187的表达水平.在MDA-MB-231细胞中分别转染miR-187 mimics(m...     

毛蕊花苷通过miR-708-3p抑制乳腺癌MCF-7细胞的恶性表型

目的 探究毛蕊花苷对乳腺癌MCF-7细胞增殖、迁移、侵袭及上皮-间质转化(EMT)的影响.方法 人乳腺癌MCF-7细胞随机分为对照组(生理盐水)以及实验组(毛蕊花苷50μmol·L-1).给药24 h后,CCK-8法检测各组细胞存活率;Transwell实验检测各组细胞迁移和侵袭能力;Western blot检测EMT...     

姜黄素诱导乳腺癌MCF-7细胞凋亡

目的 研究姜黄素对人乳腺癌细胞株MCF-7细胞增殖抑制和诱导凋亡作用.方法 MTT法检测姜黄素对MCF-7细胞的增殖抑制作用;流式细胞术(FCM)PI单染检测细胞周期;Annexin V/PI双染法检测细胞凋亡;Western blot法检测Bcl-2和Bax蛋白的表达.结果 姜黄素对MCF-7细胞生长有明显抑制作用,并呈剂量、时间依赖性;姜黄素能使MCF-7细胞阻滞在G1/S期,可以诱导细胞凋亡,Bax蛋白表达上调,而Bcl-2的表达减少.结论 姜黄素对人乳腺癌MCF-7细胞的增殖具有显著的抑制作用并可诱导细胞凋亡.其分子作用机制可能与其上调Bax基因表达水平的同时下调Bcl-2基因表达水平,从而诱导细胞凋亡有关.     

Curcumin sensitizes prostate cancer cells to radiation partly via epigenetic activation of miR-143 and miR-143 mediated autophagy inhibition

Curcumin has been reported as a radiosensitizer in prostate cancer. But the underlying mechanism is not well understood. In this study, we firstly assessed how curcumin affects the expression of miR-143/miR-145 cluster. Then, we investigated whether miR-143 is involved in regulation of radiosensitivity and its association with autophagy in prostate cancer cells. Our data showed that PC3, DU145 and LNCaP cells treated with curcumin had significantly restored miR-143 and miR-145 expression. Curcumin showed similar effect as 5-AZA-dC on reducing methylation of CpG dinucleotides in miR-143 promoter. In addition, curcumin treatment reduced the expression of DNMT1 and DNMT3B, which contribute to promoter hypermethylation of the miR-143/miR-145 cluster. Therefore, we infer that curcumin can restore miR-143 and miR-145 expression via hypomethylation. MiR-143 overexpression and curcumin pretreatment enhanced radiation induced cancer cell growth inhibition and apoptosis. MiR-143 and curcumin remarkably reduced radiation-induced autophagy in PC3 and DU145 cells. MiR-143 overexpression alone also reduced the basal level of autophagy in DU145 cells. Mechanistically, miR-143 can suppress autophagy in prostate cancer cells at least via downregulating ATG2B. Based on these findings, we infer that curcumin sensitizes prostate cancer cells to radiation partly via epigenetic activation of miR-143 and miR-143 mediated autophagy inhibition.     

LINC00094靶向miR-19b对非小细胞肺癌增殖、侵袭的抑制作用及其分子机制

目的 探讨LINC00094对非小细胞肺癌A549细胞增殖、凋亡、迁移、侵袭的影响及其作用机制。方法 将pcDNA3.1、pcDNA3.1-LINC00094、si-NC、si-LINC00094转染至非小细胞肺癌A549细胞中;将pcDNA3.1-LINC00094分别与miR-NC、miR-19b转染至A549细胞中,分别记为pcDNA3.1-LINC00094+miR-NC组、pcDNA3.1-LINC00094+miR-19b组。采用实时荧光定量PCR检测LINC00094和miR-19b表达水平;采用蛋白质印迹法检测细胞周期蛋白和凋亡蛋白表达水平;采用四甲基偶氮唑盐比色法、流式细胞术和Transwell分别检测细胞增殖、凋亡、迁移和侵袭能力;双荧光素酶报告实验检测LINC00094与miR-19b的靶向关系。结果 LINC00094在肺癌组织中低表达。过表达LINC00094可降低A549细胞的生存率,并抑制细胞迁移、侵袭（P<0.05）,且伴有相关蛋白表达水平的改变（P<0.05）。LINC00094靶向调控miR-19b,过表达miR-19b可逆转LINC00...     

Targeting miR-205 in breast cancer

As small non-coding regulatory RNAs, microRNAs are capable of silencing gene expression by translational repression or mRNA degradation. Accumulating evidence indicates that deregulation of microRNAs is often associated with human malignancies and suggests a causal role of microRNAs in neoplasia, presumably because microRNAs can function as oncogenes or tumor suppressors. Among them, miR-205 is significantly underexpressed in breast tumors compared with matched normal breast tissue although miR-205 has been shown to be upregulated in some other type of tumors. Furthermore, breast cancer cell lines, including MCF-7 and MDA-MB-231, express a lower level of miR-205 than the non-malignant MCF-10A cells. Ectopic expression of miR-205 significantly inhibits cell proliferation and anchorage-independent growth as well as cell invasion. These findings establish the tumor suppressive role of miR-205, which is probably through direct targeting of oncogenes such as ErbB3 and Zeb1. Therefore, miR-205 may serve as a unique therapeutic target for breast cancer.