Long Noncoding RNA UCA1 Targets miR-122 to Promote Proliferation,Migration, and Invasion of Glioma Cells

Glioma is the most common and lethal malignant intracranial tumor. Long noncoding RNAs (lncRNAs) have been identified as pivotal regulators in the tumorigenesis of glioma. However, the role of lncRNA urothelial carcinoma-associated 1 (UCA1) in glioma genesis is still unknown. The purpose of this study was to investigate the underlying function of UCA1 on glioma genesis. The results demonstrated that UCA1 was upregulated in glioma tissue and indicated a poor prognosis. UCA1 knockdown induced by si-UCA1 significantly suppressed the proliferative, migrative, and invasive activities of glioma cell lines (U87 and U251). Bioinformatics analysis and luciferase reporter assay verified the complementary binding within UCA1 and miR-122 at the 3¢-UTR. Functional experiments revealed that UCA1 acted as an miR-122 “sponge” to modulate glioma cell proliferation, migration, and invasion via downregulation of miR-122. Overall, the present study demonstrated that lncRNA UCA1 acts as an endogenous sponge of miR-122 to promote glioma cell proliferation, migration, and invasion, which provides a novel insight and therapeutic target in the tumorigenesis of glioma.     

miR-155 increases stemness and decitabine resistance in triple-negative breast cancer cells by inhibiting TSPAN5

Effective therapeutic targets for triple-negative breast cancer (TNBC), a special type of breast cancer (BC) with rapid metastasis and poor prognosis, are lacking, especially for patients with chemotherapy resistance. Decitabine (DCA) is a Food and Drug Administration-approved DNA methyltransferase inhibitor that has been proven effective for the treatment of tumors. However, its antitumor effect in cancer cells is limited by multidrug resistance. Cancer stem cells (CSCs), which are thought to act as seeds during tumor formation, regulate tumorigenesis, metastasis, and drug resistance through complex signaling. Our previous study found that miR-155 is upregulated in BC, but whether and how miR-155 regulates DCA resistance is unclear. In this study, we demonstrated that miR-155 was upregulated in CD24⁻CD44⁺ BC stem cells (BCSCs). In addition, the overexpression of miR-155 increased the number of CD24⁻CD44⁺ CSCs, DCA resistance and tumor clone formation in MDA-231 and BT-549 BC cells, and knockdown of miR-155 inhibited DCA resistance and stemness in BCSCs in vitro. Moreover, miR-155 induced stemness and DCA resistance by inhibiting the direct target gene tetraspanin-5 (TSPAN5). We further confirmed that overexpression of TSPAN5 abrogated the effect of miR-155 in promoting stemness and DCA resistance in BC cells. Our data show that miR-155 increases stemness and DCA resistance in BC cells by targeting TSPAN5. These data provide a therapeutic strategy and mechanistic basis for future possible clinical applications targeting the miR-155/TSPAN5 signaling axis in the treatment of TNBC.     

MiR-92b-3p ameliorates inflammation and autophagy by targeting TRAF3 and suppressing MKK3-p38 pathway in caerulein-induced AR42J cells

Acute pancreatitis (AP) is an inflammatory disease with high morbidity and mortality. Dysregulation of microRNAs (miRNAs) was involved in human diseases, including AP. However, the effects of miR-92b-3p on AP process and its mechanism remain not been fully clarified. The expression levels of miR-92b-3p and tumor necrosis factor receptor-associated factor-3 (TRAF3) were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of TRAF3, tumor necrosis factor α (TNF-α) TNF-α, interleukin-6 (IL-6), phosphorylated mitogen-activated protein kinase kinase 3 (p-MKK3), MKK3, p38 and phosphorylated p38 (p-p38) were detected by western blot. The concentration of TNF-α and IL-6 in the medium was measured using ELISA kits. The possible binding sites of miR-92b-3p and TRAF3 were predicted by TargetScan and verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The expression level of miR-92b-3p was decreased and TRAF3 expression was increased in AR42J cells stimulated with caerulein. Moreover, the protein levels of pro-inflammatory cytokines (TNF-α and IL-6) were markedly elevated, and the expression levels of autophagy-related markers Beclin1 as well as the ratio of LC3-II/I were obviously increased in AR42J cells treated with caerulein. In addition, overexpression of miR-92b-3p or knockdown of TRAF3 significantly suppressed the release of pro-inflammatory cytokines and autophagy in caerulein-induced AR42J cells. Furthermore, TRAF3 was a direct target of miR-92b-3p and its upregulation reversed the effects of miR-92b-3p overexpression on inflammatory response and autophagy. Besides, overexpression of miR-92b-3p inhibited the activation of the MKK3-p38 pathway by affecting TRAF3 expression. In conclusion, miR-92b-3p attenuated inflammatory response and autophagy by downregulating TRAF3 and suppressing MKK3-p38 pathway in caerulein-induced AR42J cells, providing a novel avenue for treatment of AP.     

利用耳甲腔型小耳畸形残耳皮瓣及软骨改善再造耳颅耳沟效果观察

目的探讨对先天性耳甲腔型小耳畸形患者行全扩张法全耳再造术后，利用残耳皮瓣改善再造耳颅耳沟的效果。方法回顾分析 2012 年 1 月—2017 年 1 月收治的 150 例先天性耳甲腔型小耳畸形患者。其中男 92 例，女 58 例；年龄 6.5～35.0 岁，平均 11.1 岁。采用一期扩张器埋置、二期全扩张法全耳再造术后发现上部颅耳沟浅显；于 6～12 个月后行三期再造耳修整。将残耳垂通过“Z”字改型转移以再造耳垂。在残耳上部作蒂在轮屏切迹的残耳上部皮瓣，弧形切开松解并加深上部颅耳沟，将上部残耳皮瓣向颅耳沟创面旋转推进缝合以覆盖创面；将带皮下组织蒂的残耳软骨组织瓣插入支架底部形成的腔隙内，并缝合固定，以增加支架的高度；耳甲腔区其余残耳皮瓣用以覆盖耳甲腔创面。结果术后拆线时 1 例患儿皮瓣远端出现直径约 0.5 cm 的表皮水疱，经换药 2 周后愈合；其余患者皮瓣成活良好。患者均获随访，随访时间 6～12 个月，平均 9.6 个月。再造耳上部颅耳沟均明显加深，再造耳支架高度不同程度增加，双耳对称性佳，耳甲腔无明显挛缩变小，再造耳外观满意。再造耳上部表面毛发明显减少，耳周发际线上移。结论采用耳甲腔型小耳畸形的残耳皮瓣及残耳软骨瓣转移，不仅可加深颅耳沟，而且可增加上部支架的高度，术后颅耳沟变形较轻，再造耳与正常耳廓的对称性更佳。     

miR-601靶向调控MMP-17抑制非小细胞肺癌的迁移和侵袭CSCD

目的探讨miR-601对非小细胞肺癌(NSCLC)迁移和侵袭能力的影响并探讨其可能的作用机制。方法RT-qPCR检测NSCLC组织中miR-601的表达,并分析表达水平与癌细胞侵袭和淋巴结转移的相关性。将miR-601 mimics瞬时转染A549细胞或H1299细胞,Transwell实验检测其对两种细胞迁移和侵袭能力的影响。生物信息学预测miR-601相关靶基因,采用双荧光素酶实验进行靶基因验证。检测miR-601的靶基因对A549细胞或H1299细胞迁移及侵袭能力的影响。结果miR-601在NSCLC组织中表达明显降低(P<0.001),且降低水平与癌细胞的浸润(P<0.007)及淋巴结转移(P<0.011)密切相关。瞬时转染miR-601 mimics能明显抑制A549细胞或H1299细胞的迁移及侵袭能力。生物信息学及荧光素酶报告实验提示MMP-17是miR-601的靶基因。MMP-17能促进A549细胞或H1299细胞的迁移和侵袭,但其促进迁移侵袭的能力可被miR-601抑制。结论miR-601通过靶向调控MMP-17而抑制非小细胞肺癌的迁移和侵袭。     

靶向PSCA的CAR-NK-92细胞对宫颈癌的抗瘤作用

目的：构建并验证靶向前列腺干细胞抗原（prostate stem cell antigen，PSCA）的嵌合抗原受体（chimeric antigen receptor，CAR）修饰的NK-92细胞对宫颈癌的抗瘤活性。方法：构建PSCA靶向的CAR慢病毒表达载体，利用慢病毒感染法获得PSCA CAR-NK-92细胞。用流式细胞术及Western blotting实验检测宫颈癌细胞中PSCA的表达水平。通过体外效靶细胞共孵育实验以及体内裸鼠移植瘤模型中的治疗情况，验证PSCA CAR-NK-92细胞对宫颈癌Hela及MS751细胞杀伤能力及其裸鼠移植瘤生长的抑制能力。结果：成功构建 PSCA CAR-NK-92 细胞，PSCA 在宫颈癌细胞中高表达（均 P<0.01）。体外共孵育结果显示，PSCA CAR-NK-92细胞可以以剂量依赖的方式裂解PSCA+的宫颈癌细胞；体内抗肿瘤数据表明，PSCA CAR-NK-92细胞较转染空载体的NK-92细胞显著抑制宫颈癌移植瘤的生长（P<0.01），并且可以有效地浸润肿瘤组织并高水平分泌TNF-α和IFN-（γ 均P<0.01）。结论：靶向PSCA的CAR-NK-92细胞在体内外都显示出了良好的对PSCA+肿瘤细胞的杀伤能力，有潜力成为一种针对宫颈癌的潜在治疗策略。     

miR-34c-3p与M2型肿瘤相关巨噬细胞在三阴性乳腺癌中的表达及意义

目的:检测miR-34c-3p与M2型肿瘤相关巨噬细胞（tumor-associated macrophage,TAM)在三阴性乳腺癌(triple negative breast cancer,TNBC)中的表达，并探讨其在乳腺癌发病中的意义。方法:选择2017年2月至2018年1月于西京医院就诊的68例TNBC患者作为实验组(A组)，以同期就诊的70例乳腺纤维腺瘤患者作为对照组(B组)。采用Real-time RT-PCR 检测组织标本中miR-34c-3p的表达；流式细胞检测M2型TAM的表达；ELISA法检测血清中IL-10的表达。并进一步分析miR-34c-3p的表达与乳腺癌M2型TAM及血清IL-10水平的相关性。结果:A、B组miR-34c-3p的相对表达量分别为0.84±0.08、1.29±0.71，A组明显低于B组，差异有统计学意义(P<0.05)。与B组比较，A组M2型TAM(CD68+CD163+)细胞比例显著升高(P<0.05)。A组血清IL-10的表达［(19.69±4.93) pg/ml］较B组［(17.26±3.51) pg/ml］显著升高，差异有统计学意义(P<0.05)。TNBC组织中miR-34c-3p的表达与M2型TAM比例及血清IL-10表达均呈显著负相关(r=-0.508，r=-0.656，P<0.05)。结论:TNBC组织中miR-34c-3p的表达下调，M2型TAM比例及血清IL-10表达均显著升高，促进TNBC进展。     

miR-96与miR-103在肝癌组织中的表达及其与预后的关系

目的 探讨miR-96与miR-103在肝癌组织中的表达及其与预后的关系。方法 选取122例肝细胞癌患者作为本研究对象。聚合酶链式反应检测miR-96和miR-103表达水平，采用单因素和多因素Cox回归模型，探讨肝癌患者中位生存时间的危险因素并建立预测模型。结果 肝癌组织中miR-96和miR-103相对表达水平均显著高于癌旁正常组织；肝癌组织中miR-96和miR-103的表达与肿瘤直径、淋巴结转移、临床分期及分化程度均有关（均P<0.05），而与性别、年龄及合并肝硬化等无关（P>0.05）；所有患者随访60月，中位生存时间为37.8月。单因素分析结果显示：肝细胞癌患者中位生存时间与肿瘤直径、淋巴结转移、TNM分期、病理分化程度、miR-96和miR-103表达水平差异均有统计学意义（均P<0.05）。多因素分析结果显示：TNM分期为Ⅲ~Ⅳ、低分化、淋巴结转移、miR-96和miR-103高表达是肝癌患者中位生存时间独立危险因素。结论 肝癌组织中miR-96和miR-103的高表达是患者预后差的独立影响因素，可作为原发性肝癌患者预后评估指标。     

肿瘤相关巨噬细胞相关性miR-99a对子宫内膜癌细胞生长和侵袭的调控作用

背景与目的：肿瘤相关巨噬细胞（tumor-associated macrophage，TAM）浸润与肿瘤进展密切相关，但作用机制尚不明确，因此，探索miR-99a对单核巨噬细胞极化的影响及其对子宫内膜癌（endometrial cancer，EC）细胞生长、侵袭的影响。方法：检测EC组织中巨噬唾液酸蛋白（macrosialin）CD68表达并分析其与临床病理学特征之间的关系；运用人EC细胞系HEC-1B、RL95-2培养上清液诱导人单核细胞U937向TAM（M2型巨噬细胞）分化；将人工合成的miR-99a模拟物片段转染至诱导后的巨噬细胞，转染后运用实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）及流式细胞术检测巨噬细胞相关因子CD68、CD163以及巨噬细胞甘露糖受体（mcrophage mannose receptor）CD206表达量变化，并运用酶联免疫吸附法（enzyme-linked immunosorbent assay，ELISA）检测巨噬细胞分泌相关细胞因子IL-12、IL-4和IL-10分泌量变化；将转染miR-99a的诱导后巨噬细胞与EC细胞共培养，运用细胞计数试剂盒（cell counting kit-8，CCK-8）和transwell法检测其对EC细胞增殖和侵袭能力的影响，并初步分析其可能的作用机制。结果：EC组织CD68高表达并与肿瘤肌层浸润及血管生成呈正相关；肿瘤细胞培养上清液成功诱导单核细胞向M2型TAM极化。转染miR-99a后单核细胞组CD68及CD163表达较对照组下降（P＜0.01），而CD206表达差异无统计学意义（P＞0.05），流式细胞术进一步证实上述表达变化；ELISA结果发现，转染miR-99a诱导后巨噬细胞中IL-12分泌增多（P＜0.01），而IL-4、IL-10分泌减少（P＜0.01），提示巨噬细胞向M2型极化受抑制。将诱导后巨噬细胞与EC细胞共培养，共培养后EC细胞增殖侵袭能力较对照组增加，而转染miR-99a模拟片段至诱导后巨噬细胞能够抑制其对增殖（P＜0.01）及侵袭能力的促进作用（P＜0.05）。诱导后巨噬细胞中过表达miR-99a后细胞中mTOR及其通路受到抑制。结论：EC间质巨噬细胞浸润与肿瘤肌层浸润及血管新生相关，miR-99a能够逆转单核细胞向M2表型极化，并抑制EC细胞介导TAM的促EC细胞生长和侵袭作用，其作用可能通过调控mTOR通路产生。