MEG3在肿瘤中作用的研究进展

长链非编码RNA(lncRNAs)自从被发现可能参与细胞生物进程以来便引起热切关注,某些lncRNA参与肿瘤发生、发展的生物进程,如INK4基因座反义非编码RNA、HOX反义基因间RNA、牛磺酸上调基因1等,而某些则表现出具有抗肿瘤的能力,如生长停滞特异性转录本5、母系表达基因(MEG3)等。MEG3作为具有抗肿瘤特性的lncRNAs中代表性的一员,成为研究的热点,MEG3表达的缺失参与多种肿瘤的发生,并且其低表达水平与基因启动子的甲基化水平显著相关。MEG3抗肿瘤的作用可能存在多种机制,如调节p53通路、作为竞争性内源性RNA调节靶基因表达等。     

lncRNA MEG3经miR-421调节E-cadherin抑制乳腺癌细胞EMT

目的探讨lncRNA母系表达基因3(MEG3)经miR-421调节E-上皮钙黏附素(E-cadherin)抑制乳腺癌细胞上皮间充质转化(EMT)的机制。 方法qRT-PCR法检测45例乳腺癌及癌旁组织lncRNA MEG3 mRNA表达。通过生物信息学网站筛选靶向lncRNA MEG3的miRNA。乳腺癌MCF-7细胞分别转染miR-NC、lncRNA MEG3 RNAi和pCDNA3.0-HA-lncRNA MEG3质粒,qRT-PCR检测各组细胞lncRNA MEG3、miR-421和E-cadherin mRNA表达水平;MTT法、Transwell法检测细胞增殖和迁移情况。 结果乳腺癌组织lncRNA MEG3表达低于癌旁组织(P＜0.05)。lncRNA MEG3与miR-421存在靶向调节作用。与阴性对照组比较,细胞lncRNA MEG3和E-cadherin mRNA表达在MEG3 RNAi组降低,MEG3质粒组增加;miR-421表达、细胞增殖率和迁移数在MEG3 RNAi组增加,MEG3质粒组降低(P＜0.05)。 结论lncRNA MEG3通过miR-421靶向调控E-cadherin,抑制乳腺癌细胞EMT。     

LncRNA MEG3对卵巢上皮癌细胞自噬的调控作用

目的探讨长链非编码RNA母系表达基因3(Lnc RNA MEG3)在卵巢上皮癌自噬中的调控机制。方法选择2016年2月-2017年7月在宜昌市中心人民医院确诊为卵巢上皮癌的患者,行卵巢切除术分别取正常卵巢组织和肿瘤组织样本。体外培养SK-OV-3细胞,对照组转染空质粒,实验组转染过表达Lnc RNA MEG3质粒;q PCR检测基因表达,蛋白印迹检测蛋白表达。结果相对于正常组织,卵巢上皮癌组织Lnc RNA MEG3表达明显降低(t=4.838,P=0.000 7),自噬相关因子LC3-Ⅱ表达明显升高(t=4.707,P=0.000 8),Lnc RNA MEG3与LC3-Ⅱ表达呈负相关(R2=0.767 5,P=0.022 1)。SK-OV-3细胞转染后,实验组比对照组Lnc RNA MEG3表达水平明显升高,差异有统计学意义(t=3.212,P=0.032 5)。实验组比对照组LC3-Ⅱ蛋白表达降低,差异有统计学意义(t=3.456,P=0.012 2)。与对照组相比,LC3-Ⅱ/LC3-Ⅰ比值也下降,差异有统计学意义(t=3.500,P=0.024 9)。结论在卵巢上皮癌中,Lnc RNA MEG3可通过负性调控激活细胞自噬从而抑制肿瘤。     

LncRNA MEG3对口腔鳞状细胞癌细胞增殖、侵袭及凋亡的影响

目的 本研究旨在探究LncRNA MEG3在口腔鳞状细胞癌（oral squamous cell carcinoma, OSCC）组织和细胞中的表达水平及对OSCC细胞增殖、侵袭和凋亡的影响。 方法 qRT-PCR法检测OSCC患者口腔粘膜组织、肿瘤组织、及Tca8113细胞系中LncRNA MEG3的表达水平。构建pcDNA3.1-MEG3重组质粒转染Tca8113细胞,分为3组：转染过表达pcDNA3.1-MEG3的质粒（MEG3组）和空质粒pcDNA3.1-NC （NC组）;空白对照组（BLANK组）加入PBS。分别采用qRT-PCR法、CCK-8法、流式细胞术和Transwell检测LncRNA MEG3表达、细胞增殖、凋亡率和侵袭能力。结果 与正常口腔粘膜组织和细胞相比,LncRNA MEG3在OSCC组织和细胞中的表达下调（P<0.05）。体外实验发现,与空白对照组和NC组相比,过表达LncRNA MEG3组细胞增殖和侵袭能力受到抑制,凋亡率明显升高（P<0.05）。结论 LncRNA MEG3在OSCC组织和细胞中低表达;LncRNA MEG3过表达可抑制OCSS细胞的增殖和侵袭,促进凋亡。     

长链非编码RNA母系印记基因3对人类绒毛发育的作用

目的 研究长链非编码RNA (lncRNA)母系印记基因3(MEG3)对人类流产绒毛发育的影响,并探讨其分子机制.方法 收集自然流产及人工流产各15例患者的绒毛组织,使用免疫组化方法检测其凋亡因子Bax及凋亡抑制因子Bcl-2的表达,使用qPCR法检测MEG3的表达.在人滋养细胞系HTR-8/SVneo细胞中过表达MEG3后,利用qPCR法检测MEG3的表达,利用细胞侵袭实验比较过表达组与对照组细胞的侵袭能力.结果 人工流产绒毛组织中Bax的表达低于自然流产绒毛,而Bcl-2的表达高于自然流产绒毛(P＜0.01).人工流产绒毛组织中的MEG3的表达低于自然流产(P＜0.01).人滋养层细胞系HTR-8/SVneo细胞中过表达MEG3后,HTR-8/SVneo细胞侵袭能力降低(P＜0.01).结论 IncRNA MEG3可调控滋养层细胞的凋亡及侵袭能力,影响人类绒毛的发育.     

长链非编码RNA MEG3对胃癌细胞增殖的影响研究

目的:探讨长链非编码RNA MEG3在胃癌组织及细胞系中的表达水平,以及过表达MEG3对胃癌细胞增殖能力的影响,并探索其可能的作用机制。方法:用定量PCR（qRT-PCR）技术检测胃癌组织及细胞系中MEG3的表达水平;通过转染pCDNA-MEG3上调MEG3的表达水平,并通过qRT-PCR检测转染效率。用MTT和克隆形成实验检测上调MEG3的水平对BGC-823细胞和MGC-803细胞的增殖能力的影响,用Western blot实验检测这两种细胞中上调MEG3的水平对p53蛋白的表达水平的影响。结果:相比正常胃组织及细胞,在胃癌组织和细胞中MEG3的表达出现显著下调,SGC7901、MGC-803和BGC-823细胞中MEG3的表达水平分别是正常胃上皮细胞GES-1中的73.6 %、42.0 %和27.1 % （p<0.05）。BGC-823细胞和MGC-803细胞中转染pCDNA-MEG3能显著上调MEG3的表达,MEG3的表达水平分别为对照组的252倍和311倍（p<0.05）;MTT和克隆形成实验显示,上调MEG3的表达能降低BGC-823细胞和MGC-803细胞的增殖能力。Western blot实验显示,转染了pCDNA-MEG3的MGC-803和BGC-823细胞中p53的表达相较对照组中显著增加。结论:胃癌组织及细胞中MEG3的表达下调,且这可能通过抑制p53蛋白的激活从而促进胃癌细胞增殖,影响胃癌的发生和发展。     

最小努力(惰性)原则与慢性病预防行为选择

目的:探讨长链非编码RNA MEG3在胃癌组织及细胞系中的表达水平,以及过表达MEG3对胃癌细胞增殖能力的影响,并探索其可能的作用机制?方法:用定量反转录PCR(qRT-PCR)技术检测胃癌组织及细胞系中MEG3的表达水平;通过转染pcDNA-MEG3上调MEG3的表达水平,并通过qRT-PCR检测转染效率?用MTT和克隆形成试验检测上调MEG3的水平对BGC-823细胞和MGC-803细胞的增殖能力的影响,Western blot检测这两种细胞中上调MEG3的水平对p53蛋白的表达水平的影响?结果:相比正常胃组织及细胞,在胃癌组织和细胞中MEG3的表达出现显著下调,SGC7901?MGC-803和BGC-823细胞中MEG3的表达水平分别是正常胃上皮细胞GES-1中的73.6%?42.0%和27.1%(P < 0.05)?BGC-823细胞和MGC-803细胞中转染pcDNA-MEG3能显著上调MEG3的表达,MEG3的表达水平分别为对照组的252倍和311倍(P < 0.05);MTT和克隆形成试验显示,上调MEG3的表达能降低BGC-823细胞和MGC-803细胞的增殖能力?Western blot实验显示,转染了pcDNA-MEG3的MGC-803和BGC-823细胞中p53的表达相较对照组中显著增加?结论:胃癌组织及细胞中MEG3的表达下调,且这可能通过抑制p53蛋白的激活从而促进胃癌细胞增殖,影响胃癌的发生和发展?     

H3K27me3聚集MEG3 lncRNA启动子通过MDM2/p53途径诱导多发性骨髓瘤RPMI8226细胞凋亡逃逸

目的 从组蛋白甲基化与长链非编码 RNA ( ln-cRNA)相互作用角度探讨多发性骨髓瘤( MM)细胞凋亡逃逸的具体机制.方法 MTT法检测细胞增殖活性;Annex-inV-FITC/PI双标流式细胞术检测下调组蛋白H3第27位赖氨酸上三甲基化H3K27me3 的表达是否可以诱导多发性骨髓瘤细胞株—RPMI8226 细胞凋亡; RT-PCR 法检测 RP-MI8226细胞中人母系表达基因( MEG3) lncRNA及鼠双微体基因 2 ( MDM2 )的 mRNA 表达; Western blot 检测 RP-MI8226 细胞中 Zeste 基因增强子同源物 2 ( EZH2 )、H3K27me3、MDM2 及 p53 蛋白表达; ChIP Real -time PCR检测RPMI8226细胞中 H3K27me3 的结合位点.结果 在RPMI8226 细胞中,下调 H3K27me3 可诱导细胞凋亡;H3K27me3可直接结合于 MEG3lncRNA 启动子; RPMI8226细胞中抑制了 H3K27me3 可上调 MEG3lncRNA 表达; RP-MI8226经MEG3lncRNA 小干扰 RNA ( MEG3lncRNA -siR-NA)干预下调MEG3lncRNA时,细胞内MDM2 mRNA水平明显升高,并诱导 MDM2 蛋白表达.给予 MDM2 拮抗剂后, Ub-p53 表达降低, p53 降解被抑制.结论 MM 中H3K27me3结合MEG3 lncRNA启动子,MEG3lncRNA启动子H3K27me3高表达导致MEG3 lncRNA表达缺失. MEG3 ln-cRNA表达缺失解除MDM2的抑制, MDM2与p53直接结合介导p53泛素化,促使p53降解,细胞凋亡逃逸.     

姜黄素调控LncRNA MEG3/miR-21/PTEN通路促HPV阳性宫颈癌细胞凋亡

目的：探究姜黄素杀伤HPV阳性宫颈癌细胞的作用机制。方法：采用细胞活性检测实验和细胞凋亡检测实验分析姜黄素对HPV阳性宫颈癌细胞活性以及细胞凋亡的影响;采用实时定量PCR实验和蛋白质印迹（Western blot）法检测姜黄素促进HPV阳性宫颈癌细胞凋亡过程中关键信号通路因子LncRNA MEG3、miR-21、PTEN和活化的caspase3的变化水平。结果：姜黄素显著促进了HPV阳性宫颈癌细胞凋亡,升高了LncRNA MEG3、PTEN和活化的caspase3的表达水平,下调了miR-21的表达（P＜0.05）;利用siRNA敲低LncRNA MEG3的表达,能够逆转姜黄素对miR-21、PTEN和caspase3的调控（P＜0.05）。结论：姜黄素可能通过上调LncRNA MEG3而降低miR-21的表达,从而上调PTEN的表达,最终促进HPV阳性宫颈癌细胞的凋亡。     

DNMT1蛋白通过沉默MEG3基因促进视网膜母细胞瘤增殖

目的 探讨 DNMT1 蛋白是否通过沉默 MEG3 基因诱导视网膜母细胞瘤增殖。方法 通过转染 pcDNA-DNMT1 或si-DNMT1上调或干扰DNMT1的表达水平;通过转染pcDNA-MEG3或si-MEG3上调或干扰MEG3的表达水平;用Western blot检测视网膜母细胞瘤细胞系中DNMT1蛋白表达量;用CCK-8法及EdU法检测细胞的增殖能力;用实时荧光定量PCR技术检测转染后的视网膜母细胞瘤细胞中MEG3表达量的变化;干扰DNMT1表达后,用甲基化特异性PCR检测MEG3基因启动子DNA甲基化水平的变化。结果 视网膜母细胞瘤SO-RB50及HXO-RB44细胞中DNMT1蛋白表达量显著升高（P<0.05）。转染了 pcDNA-DNMT1 的 HXO-RB44 细胞中 DNMT1 蛋白表达增加,细胞增殖能力增加,MEG3 表达量降低;转染了 siRNADNMT1的SO-RB50细胞DNMT1蛋白表达减少,细胞增殖能力降低,MEG3表达量增加（P<0.05）。干扰DNMT1蛋白表达后,MEG3基因启动子DNA甲基化水平降低（P<0.05）。逆转DNMT1蛋白对MEG3基因的调控后,DNMT1蛋白调控RB细胞增殖的能力减弱（P<0.05）。结论 在视网膜母细胞瘤细胞中,DNMT1蛋白表达的上调,诱导了MEG3基因启动子DNA甲基化失活,最终导致细胞异常增殖。     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

A preliminary screening study on the associated proteins in human psoriasis vulgaris by serum proteomics technologies

Objective：To investigate the optimum screening conditions of associated proteins in human psoriasis vulgaris by serum proteomics technique, and to screen the different expression proteins related with psoriasis vulgaris. Methods：Serum samples of peripheral blood were collected from newly diagnosed psoriasis vulgaris patients in the clinic, and 20 matched healthy persons. Serum albumin IgG was removed by filtering with ProteoExtract Albumin/IgG. After comparative proteomics analysis the different protein spots were identified using 2-DE and MS. Results ：Electrophoresis figures with high resolution and reproducibility were obtained. Three different expression proteins were found only in the serum from psoriasis vulgaris patients, while nine other different proteins expressing from healthy volunteers. Conclusion：The protein expression was different in the serum between the psoriasis vulgaris patients and healthy volunteers. It was hoped that we could find the biomarkers related to psoriasis vulgaris by using proteomics.     

A CASE OF HUGE GASTRIC DISOPYROBEZOAR

PYTOBEZOARS are common bezoars in gastrontestinal tract, commonly seen in stomach and mall intestine,1but huge disopyrobezoars are     

Acupuncture Treatment of Chest Bi Syndrome

CASE HISTORY A male,a retired officer of 61 years old,paid his first visit on May 12,2006.The patient stated that 2 weeks before he suddenly got paroxysmal precordial choking pain,with irregular attacks.Each attack would last several seconds to several minutes.The attack was accompanied with heavy oppressed sensation in the precordial region,palpitation,lassitude,and in severe cases,sweating,which was once diagnosed by a hospital as'coronary heart disease'.Recalling the case history,the patient said that several days before he got angry with others because of some affairs in work,and he was not so used to the life pattern since he had just retired from his busy working post.At usual times,he was quick tempered and peevish.In recent days,he had poor appetite,and loose stool.     

Instructions to Contributors

The English edition of Chinese Journal of Integrative Medicine is edited and published by the Chinese Journal of Integrated Traditional and Western Medicine Press.Original scientific papers of advanced clinical and experimental medicine in the field of complementary/alternative medicine or integrative medicine are expected.     

Development Is A High Level Inheritance for China Time-honored Brand Products

<正>As culture is the soul of a nation,the three books,"History of Chinese Philosophy,""History of Chinese Culture"and"History of Chinese Science and Technology"have demonstrated the superiority of our great Chinese culture,and its significant impact in the field of international culture.Both core value and practical value of the Chinese culture are very clear,reflected in the views of universe,nature,life,health,disease,and therapy and related considerations,concepts,and technical methods.