Abstract Substitutions at the nodal and anti-nodal positions of conjugated aromatics exert profound effects on their electronic properties, yet a systematic investigation is lacking on how these underlying quantum mechanical rules are manifested experimentally. Here, using polycyclic aromatic hydrocarbons derivatives as a model system, we systematically elucidate how nodal and anti-nodal substitutions dictate their chemical reactivity and physical properties. It is found that Sonogashira C–C coupling at the nodal position has a noticeably lower product yield than anti-nodal position under identical reflux conditions due to inhibited molecular orbital amplitude. Comprehensive spectroscopic characterization of the resulting substituted products reveals two striking photophysical differences: 1) Frontier orbital symmetry is largely conserved in node-substituted molecules, whereas the anti-nodal substitution induces strong symmetry-breaking, leading to accelerated fluorescence emission in anti-node-substituted pyrenes; 2) Nodal substitution induces the formation of charge-transfer states due to a twisted geometry, which activates room-temperature phosphorescence via improved singlet–triplet intersystem crossing. This study establishes an effective structure–property relationship linking molecular orbital symmetry to macroscopic cross-coupling reactivity and molecular photophysics within these conjugated aromatics.

Fluorination is a useful approach for tailoring the physicochemical properties of nanocarbon materials. However, owing to the violent reactivity of fluorination, achieving edge-perfluorination of nanographene while maintaining its original π-conjugated structure is challenging. Instead of using traditional fluorination, here, we employed a bottom-up strategy involving fluorine preinstallation and synthesized decafluorinated and perfluorinated warped nanographenes (DFWNG and PFWNG, respectively) through a 10-fold Suzuki-Miyaura coupling followed by a harsh Scholl reaction, whereby precisely edge-perfluorinated nanographene with an intact π-conjugated structure was achieved for the first time. X-ray crystallography confirmed the intact π-conjugated structure and more twisted saddle-shaped geometry of PFWNG compared to that of DFWNG. Dynamic study revealed that the 26-ring carbon framework of PFWNG is less flexible than that of DFWNG and the pristine WNG, enabling chirality resolution of PFWNG and facilitating the achievement of CD spectra at -10 °C. The edge-perfluorination of PFWNG resulted in improved solubility, lower lowest unoccupied molecular orbital, and a surface electrostatic potentials/dipole moment direction opposite those of the pristine WNG. Likely owing to its intact π-conjugated structure, PFWNG exhibits comparable electron mobility with well-known PC61BM. Furthermore, perfluorination improves thermal stability and hydrophobicity, making PFWNG suitable for use as a thermostable/hydrophobic n-type semiconductor material. In the future, this fluorination strategy can be used to synthesize other perfluorinated nanocarbon materials, such as perfluorinated graphene nanoribbons and porous nanocarbon.

目的 探讨CARM1与p54nrb在浸润性乳腺癌组织中的表达情况及其与临床病理学特征的关系.方法 采用免疫组织化学染色SP法检测121例浸润性乳腺癌组织及随机抽取与其配对的60例癌旁正常组织中CARM1和p54nrb的表达情况,分析CARM1和p54nrb表达与乳腺癌临床病理学特征及乳腺癌分子分型的关系.结果 CARM1在乳腺癌中的阳性表达率为55.4％,高于癌旁组织(38.3％);p54nrb的阳性表达率为57.9％,高于癌旁组织(35.0％),以上差异均具有统计学意义(P＜0.05).CARM1和p54nrb在浸润性乳腺癌中的表达水平与较高组织学分型,ER、PR、高Ki-67指数相关,与患者发病年龄、肿瘤大小、淋巴结转移(N)、TNM分期及HER-2表达情况无关(P＞0.05).CARM1表达与p54nrb表达呈正相关(r=0.463,P=0.000).结论 乳腺癌组织中CARM1与p54nrb均呈高表达,提示两者可能与乳腺癌的发生、高恶性程度及侵袭力相关.

目的 探讨β-连环蛋白(β-catenin)、染色质重塑蛋白1(BRG1)及解整合素金属蛋白酶17(ADAM17)在结肠癌组织中的表达水平及其与临床病理特征之间的关系.方法 采用免疫组化法检测70例结肠癌组织及20例癌旁组织中β-catenin、BRG1及ADAM17的表达水平,并分析其与临床病理特征之间的联系.结果 β-catenin、BRG1及ADAM17在结肠癌组织中的阳性表达率分别为62.9％、71.4％、62.9％,显著高于相应癌旁组织(25％、30％、35％),差异有统计学意义(P<0.05);β-catenin、BRG1、ADAM17阳性表达与结肠癌分化程度、T分期、临床分期和血管侵犯有关,Spearman相关性分析结果显示β-catenin、BRG1和ADAM17相互之间呈正相关.结论 β-catenin、BRG1、ADAM17在结肠癌组织中表达上调,与结肠癌的发生发展有关,可作为判断预后及靶向治疗的重要依据.

目的 通过调高MCF-7人乳腺癌细胞ADAM17的表达,探讨其对乳腺癌细胞侵袭力的影响.方法 MCF-7细胞成功转染慢病毒载体(LV8N-ADAM17)后,实验分为转染组、无义序列组、空白对照组,通过荧光显微镜观察转染效果.采用实时荧光定量PCR、蛋白质印迹法分别检测各组MCF-7细胞ADAM17、EGFRmRNA和ADAM17、EGFR蛋白的表达情况;采用细胞划痕实验与Transwell小室法分别检测过表达ADAM17对MCF-7细胞迁移力和侵袭力的影响.采用SPSS 17.0对数据进行统计学分析,组间比较采用单因素方差分析,两两多重比较采用LSD检验.结果 MCF-7细胞转染过表达的ADAM 17慢病毒,转染率为100％.转染组ADAM17mRNA表达量为1.79±0.08,高于无义序列组的1.16±0.07和空白对照组的1.13±0.05,差异有统计学意义,F=90.684,P＜0.001;转染组EGFRmR-NA表达量为1.72±0.15,高于无义序列组的1.21±0.04和空白对照组的1.17±0.09,F=26.215,P＜0.001;无义序列组和空白对照组的ADAM17mRNA、EGFRmRNA的表达量差异均无统计学意义,均P＞0.05.转染组ADAM17蛋白相对表达量为0.81±0.07,高于无义序列组0.52±0.04和空白对照组0.50±0.02,F=39.231,P＜0.001;转染组EGFR蛋白相对表达量为1.43±0.24、高于无义序列组的0.86±0.06和空白对照组的0.88±0.17,F=10.483,P＜0.001;无义序列组和空白对照组的ADAM17和EGFR蛋白表达量差异无统计学意义,均P＞0.05.转染组乳腺癌细胞划痕愈合率为(70.67±9.01)％,高于无义序列组(37.00±2.65)％和空白对照组(39.00±4.58)％,差异有统计学意义,F=85.784,P＜0.001.转染组的穿膜细胞数为(101.00±6.19)个/视野,高于无义序列组(76.35±6.41)个/视野和空白对照组(79.20±10.04)个/视野,差异有统计学意义,F=13.499,P＜0.001.结论 慢病毒载体有效地将过表达AD-AM17转染入MCF-7乳腺癌细胞,MCF-7乳腺癌细胞的侵袭力被显著提高.

三阴性乳腺癌是一种特殊的乳腺癌亚型,占所有乳腺癌类型的15％～20％,其具有高度异质性、较强侵袭力和早期易复发、转移等特点.临床上主要采取手术、放化疗的治疗方式,在早期采取新辅助化疗并进行疗效评估是目前首选的全身治疗方法.随着对分子生物学研究的不断深入,三阴性乳腺癌的分型被细化,针对不同分型制订个体化靶向治疗方案亦具有重要意义.本文就现阶段早期三阴性乳腺癌的局部治疗、新辅助化疗以及最新靶向治疗成果进行总结,以期进一步推动早期三阴性乳腺癌治疗的研究进展.

目的 探讨微RNA-145(miR-145)对三阴性乳腺癌MDA-MB-231细胞增殖、侵袭和转移能力的影响.方法 MDA-MB-231细胞分为3组,空白组(细胞无特殊处理)、miR-145 NC组(转染miR-145无义序列)、miR-145 inhibitors组(转染miR-145 inhibitors).实时聚合酶链反应检测miR-145、解聚素-金属蛋白酶17(ADAM17)和表皮生长因子受体(EGFR) mRNA的相对表达水平,CCK-8法比较各组细胞的增殖能力,Transwell实验以及划痕实验测定各组细胞的侵袭和迁移能力,Western blot检测各组ADAM17和EGFR蛋白表达情况.结果 miR-145 inhibitors组miR-145表达量低于miR-145 NC组,差异有统计学意义(P＜0.05);miR-145 NC组与空白组miR-145表达量比较,差异无统计学意义(P＞0.05).转染后24、48、72 h,空白组与miR-145 NC组光密度(OD)值比较,差异无统计学意义(P＞0.05).转染后72 h,miR-145 inhibitors组OD值大于miR-145 NC组,差异有统计学意义(P＜0.05).转染后24、48 h,miR-145 inhibitors组与miR-145 NC组OD值比较,差异无统计学意义(P＞0.05).miR-145 inhibitors组跨膜细胞数多于miR-145 NC组,差异有统计学意义(P＜0.05);miR-145 NC组与空白组跨膜细胞数比较,差异无统计学意义(P＞0.05).miR-145 inhibitors组划痕愈合率高于miR-145 NC组,差异有统计学意义(P＜ 0.05);miR-145 NC组与空白组细胞愈合率比较,差异无统计学意义(P＞0.05).miR-145 inhibitors组ADAM17 mRNA相对表达量高于miR-145 NC组,差异有统计学意义(P＜0.05);miR-145 NC组与空白组ADAM17 mRNA相对表达量比较,差异无统计学意义(P＞0.05).miR-145 inhibitors组和miR-145 NC组EGFR mRNA的相对表达量比较,差异无统计学意义(P＞0.05);miR-145 NC组与空白组EGFR mRNA相对表达量比较,差异无统计学意义(P＞0.05).miR-145 inhibitors组ADAM17、EGFR蛋白相对表达量高于miR-145 NC组,差异有统计学意义(P＜0.05);miR-145 NC组与空白组ADAM 17、EGFR蛋白相对表达量比较,差异无统计学意义(P＞0.05).结论 miR-145 inhibitor转染MDA-MB-231细胞后,通过靶向激活ADAM17/EGFR通路,促进MDA-MB-231细胞的增殖、侵袭和迁移.

Objective:To investigate the effect of a disintegrin and metalloproteinase 17 (ADAM17) - short hairpin RNA (shRNA) transfection on the proliferation of human breast cancer MCF-7 cells.Methods:Four shRNA sequences (adam17-hsa-297, adam17-hsa-1508, adam17-hsa-1658) targeting ADAM17 were designed, Adam17-hsa-1864) and a negative control (LV10-NC) were used to screen the best inhibition rate of ADAM17 shRNA by qPCR.The experiment was divided into three groups: transfection group, meaningless sequence group and control group.RNA was extracted according to routine steps, and then reverse transcripted and amplified.The expression of ADAM17 mRNA was detected by qPCR and the proliferation of MCF-7 cells was measured by MMT method.Results:The results of MTT assay showed that the absorbance values of control group, nonsense sequence group and transfection group were 0.270±0.040, 0.250±0.035 and 0.185±0.080, respectively.There was significant difference between the two groups ( F=3.854, P=0.045). There was no significant difference between the control group and the nonsignificant sequence group ( P>0.05), and the difference between the control group and the transfection group was statistically significant ( P<0.05). There was no significant difference between the nonsignificant sequence group and the transfection group ( P>0.05); the absorbance values of the control group, the meaningless sequence group and the transfection group at 48 h were 0.500±0.057, 0.494±0.086 and 0.311±0.007, respectively, and the differences between the two groups were statistically significant( F=19.42, P<0.001). There were significant differences between transfection group and control group and no significant sequence group (all P<0.05), but there was no significant difference between control group and non significant sequence group ( P>0.05). The absorbance values of control group, nonsense sequence group and transfection group at 72 h were 0.720±0.150, 0.713±0.174 and 0.558±0.071, respectively.There was no Conclusion:BMSC transfected with ADAM17 shRNA could inhibit the proliferation of MCF-7 cells at 24 h and 48 h, while the proliferation of MCF-7 cells decreased at 72 h.

Dual-phosphorescence emission can be realized via chemical conjugation between an n–π* type of luminophore and a π–π* one.

目的 MicroRNA-145(miR-145)在乳腺癌中低表达,文章通过研究miR-145与ADAM17、EGFR的关联,探究miR-145对乳腺癌MCF-7细胞的调控作用.方法 将MCF-7乳腺癌细胞分成3组,即转染组(转染miR-145 mimics)、对照组(未经转染)、无义序列组(转染无意义miRNA的无义序列).利用MTT、Transwell、实时定量荧光PCR(qPCR)技术分别检测3组MCF-7乳腺癌细胞的增殖能力、侵袭能力及转染miR-145后的表达量.利用qPCR及蛋白印迹法检测3组乳腺癌MCF-7细胞中ADAM17、EGFR mRNA和蛋白水平.结果 qPCR检测的结果提示miR-145相对表达量在转染组(13964.33±1265.30)明显高于对照组(1.00±0.05)和无义序列组(1.03±0.15),差异有统计学意义(P<0.01);转染组ADAM17 mRNA的表达量(1.71±0.08)明显高于对照组(1.00±0.07),差异有统计学意义(P<0.01).与24、48、72 h无义序列组比较,转染组MCF-7抑制率明显升高(P<0.01).Transwell侵袭实验结果显示,转染组穿膜细胞数[(56.20±2.17)个/视野]较对照组[(92.80±3.90)个/视野]、无义序列组[(91.80±4.97)个/视野]明显下降(P<0.01).蛋白印迹实验得出结果,转染组细胞中ADAM17、EGFR的蛋白含量较对照组、无义序列组显著下降(P<0.01).结论 miR-145作用于ADAM17-EGFR信号通路,从而抑制乳腺癌MCF-7细胞系增殖、侵袭.

Objective To investigate the effects of ADAM17 -shRNA on implanted tumor of MCF -7 breast cancer cells in nude mice.Methods MCF-7 breast cancer cells were transfected by lentivirus -ADAM17-shRNA. Thirty nude mice were randomly divided into transfection group(MCF-7-ADAM17-shRNA),vector group(MCF-7-shNC)and control group(MCF-7).The cell suspension were subcutaneously injected in the right side of the footpath of each nude mouse.All nude mice were sacrificed by cervical dislocation on the 26th day after implantation.The morpho-logical structure was observed by HE staining.The expression of ADAM17 and Ki-67 was assayed by IHC,and expres-sion of ADAM17 protein was assessed by Western Blot.Results When nude mice were sacrificed, the tumor sizes in transfection group,control group and vector group were 238.95 ±17.24 mm3,610.48 ±15.96 mm3and 604.50 ±15.14 mm3,respectively.There was significant difference on tumor size among the three groups(F=1 765.86, P=0.00). Compared with the transfection group,there were lager necrosis areas in vector group and control group.The positive ex-pression of ADAM17 was in the cytoplasm,and tumor staining score of ADAM17 in transfection group was 2.45 ±0.44, which was significantly lower than those in vector group(7.09 ±0.18)and control group(7.26 ±0.25)(F=163.13, P=0.00).The positive expression of Ki -67 protein was in the nuclei and the tumor staining score in transfection group was 3.54 ±0.26, which was significantly lower than those in vector group(8.92 ±0.41)and control group(9.28 ± 0.45)(F=481.95,P=0.00).The OD values of ADAM17 protein in control group,vector group and transfection group were 0.632 ±0.037,0.619 ±0.027 and 0.388 ±0.034, respectively.Compared with the vector and control group, the expression of ADAM17 protein was significantly lower in transfection group(F=56.13, P=0.00).Conclusion The growth of human breast cancer MCF -7 cells transfected with ADAM17-shRNA is significantly inhibited in nude mice.

目的 探讨利用裸鼠乳腺癌荷瘤模型转染解聚素-金属蛋白酶17 (ADAM17)-shRNA的骨髓间充质干细胞(BMSCs)对乳腺癌移植瘤的抑制效果.方法 将人乳腺癌细胞(MCF-7)接种于30支裸鼠皮下,14 d后乳腺癌荷瘤裸鼠模型成功构建.随机分为:转染组(注射1×106/ml转染ADAM17-shRNA的BMSCs)、BMSCs组(注射1×106/ml BM-SCs)和对照组(注射空白PBS).采用贴壁筛选法分离、培养3周龄雄性SD大鼠的BMSCs,利用慢病毒介导将AD-AM17-shRNA转染第3代培养的BMSCs,将其通过尾静脉注射给各组荷瘤裸小鼠(0.1 ml/次)3d重复一次,共5次,观察各组裸小鼠肿瘤的生长状况.抑瘤实验16 d后处死裸鼠,HE染色法观察各组瘤体的形态结构,免疫组化法检测各组瘤体ADAM17、表皮细胞生长因子受体(EGFR)和细胞增殖核抗原(Ki-67)的表达.结果 转染组、BMSCs组、对照组裸鼠瘤体体积分别为(375.09±20.72)、(768.85±35.55)、(783.75±27.30) mm3,转染组相较于对照组移植瘤体积缩小,差异有统计学意义(P＜ 0.05),而BMSCs组差异无统计学意义.HE染色镜下可见,与对照组相比转染组可见瘤体组织大片坏死区域、坏死区域细胞崩解、细胞结构消失,而BMSCs组与对照组差异无统计学意义.转染组ADAM17、EGFR、Ki-67蛋白与对照组相比表达均降低,差异有统计学意义(P＜0.05);BMSCs组和对照组相比较,差异无统计学意义.结论 ADAM17-shRNA通过BMSCs介导可靶向归巢至裸鼠乳腺癌移植瘤并发挥抑制裸鼠乳腺癌ADAM17、EG-FR和Ki-67表达的作用.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Objective To explore the effect of ADAM17-shRNA on implanted tumor of MCF-7 breast cancer cells in nude mice. Methods Thirty nude mice were randomly divided into transfection group (MCF-7-ADAM17-shRNA), vector group (MCF-7-shNC) and control group (MCF-7). Three groups of cell suspension were injected subcutaneously in the right side of the footpath of the corresponding nude mice. The general condition of the nude mice was observed every day. When the tumor nodules were measured, the tumor sizes were measured every 4 days. All nude mice were sacrificed by cervical dislocation on the 28th day after implantation. The morphological structures were observed after HE staining, and the expressions of ADAM17, p-EGFR, EGFR, p-AKT, AKT,p-ERK and ERK in three groups were assayed by Western blot. Results The tumor sizes of the control group, the vector group and the transfection group were (639.821 ± 15.429) mm3, (643.350 ± 15.543) mm3and (240.233 ± 10.536) mm3respectively. There were differences in tumor volume among different time points in the three groups (P < 0.05). There were differences in tumor volume among the transfection group, the control group and the vector group (P < 0.05). Compared with the transfection group, the necrosis areas in the vector and control groups were lager. The data of Western blot showed that the expressions of ADAM17, p-EGFR, EGFR, p-AKT, AKT, p-ERK and ERK in the transfection group were significantly lower than those in the control and vector groups (P < 0.05). Conclusions ADAM17-shRNA can effectively inhibit the growth of transplanted tumor of human breast cancer MCF-7 cells in nude mice. EGFR-PI3K-AKT and EGFR-MEK-ERK signaling pathways are involved in the process.

A disintegrin and metalloprotease 17 (ADAM17) is highly expressed in many malignant tumors and is closely related to their development. We showed in a previous study that silencing of ADAM17 by siRNA inhibited the growth of MCF-7 breast cancer cells in vitro and in vivo. In the present study, we investigated the effects of ADAM17-short hairpin RNA (ADAM17-shRNA) on MCF-7 breast cancer cells and explored the potential action pathway. In vitro, transfection of shRNAs was performed using a lentivirus, and the effects of ADAM17-shRNA on invasion, proliferation and cell cycle distribution of MCF-7 cells were assessed by Boyden chamber method, real-time cell analysis and flow cytometry, respectively. In vivo, MCF-7 cells with different administrations were transplanted subcutaneously into nude mice, and the effect of ADAM17-shRNA on the growth of transplanted tumors was assessed. In addition, the morphological structures were observed by H&E staining, and the expression of ADAM17 and Ki-67 was assessed by immunohistochemistry; expression of ADAM17, EGFR, p-EGFR, AKT, p-AKT, ERK and p-ERK proteins was assessed by western blotting, respectively. Our data showed that ADAM17-shRNA successfully inhibited ADAM17 mRNA expression, invasion and proliferation of MCF-7 cells resulting in G0/G1 phase arrest, and significantly inhibited the growth of transplanted tumors with larger areas of necrosis, low expression of ADAM17 and Ki-67 and reduced protein expression of ADAM17, EGFR, p-EGFR, AKT, p-AKT, ERK, and p-ERK in the tumor tissues. The present research suggests that ADAM17-shRNA can inhibit MCF-7 cell invasion and proliferation in vitro and inhibit MCF-7 xenograft growth in vivo through the EGFR/PI3K/AKT and EGFR/MEK/ERK signaling pathways.

Objective:To investigate the inhibitory effect of bone marrow mesenehymal stem cells (BMMSCs) transfected with ADAM 17-shRNA (adisintegrin and metalloprotease 17-shRNA) on the growth of implanted breast cancer MCF-7 cell xenograft in nude mice.Methods:BMMSCs from 3-week-old male SD rats were isolated and cultured with the whole bone marrow adherence method.BMMSCs were transfected with Lentivirus-mediated ADAM17-shRNA.Breast cancer MCF-7 cell xenograft model was successfully established in 30 nude mice after 14 days implantation of tumor cells.According to the random number table,nude mice were randomly divided into con trol group (equal volume PBS),BMMSCs group (1 × 106/ml BMMSCs) and transfection group (1 × 106/ml BMMSCs transfected with ADAM17-shRNA) with 10 nude mouses in each group.The tumor inhibition test was carried out on the 15th day by injecting BMMSCs into tail vein (0.1 ml/each,administration was carried every 3 days with a to tal of 5 times).The growth of implanted tumor was observed every day.All the nude mice were sacrificed on 16th day after treatment.The expressions of ADAM17 mRNA and ADAM17 protein in rumor tissues were detected by Real-time PCR and Westem blotting,respectively.Results:The volume of implanted tumor in control group,BMMSCs group was significantly larger than that of transfection group ([787.15 ±25.95],[767.02±28.98] vs [361.89±19.75] mm3,all P＜0.01) on D 30.The tumor inhibition rate of BMMSCs group and transfection group was significantly higher than that of control group (2.57％,53.89％ vs 0.00％,all P＜0.05).The expression of ADAM17 mRNA in control group,BMMSCs group was significantly higher than that of transfection group (1.00±0.01,0.97± 0.08 vs 0.30±0.09,P＜0.05).The expression of ADAM17 protein in control group,BMMSCs group was significantly higher than that of transfection group (0.70±0.09,0.68±0.02 vs 0.45±0.05,all P＜0.05).Conclusion:The tropism of ADAM17-shRNA to breast cancer xenograft in nude mice was accomplished by BMMSCs mediation,which may play an anti-tumor effect.

Objective To investigate the effect of a disintegrin and metalloprotease 17 ( ADAM17 )-shRNA on the proliferation of human breast cancer MCF-7 cells and to explore its possible mechanism .Methods MCF-7 cells were di-vided into the nonsense group , control group ( PBS) and transfection group .Four specific ADAM17-shRNAs ( ADAM17-shRNA-1219, ADAM17-shRNA-1508, ADAM17-shRNA-1134 and ADAM17-shRNA-297) were designed and synthesized for ADAM17 gene.MCF-7 cells were transfected by electroporation .The nonsense group was transfected by ADAM 17-shNC, the control group was added with PBS , and the transfection group was transfected by ADAM 17-shRNA-1219, AD-AM17-shRNA-1508, ADAM17-shRNA-1134 and ADAM17-shRNA-297, respectively.The expression of ADAM17 mRNA, the expression of ADAM17 protein, cell growth curve, proliferation activity and cell cycle of MCF-7 cells was respectively detected by real-time PCR, Western blotting, iCELLigence and flow cytometry .Results Four ADAM17-shRNA se-quences inhibited the expression of ADAM 17 gene, the shRNA1219 vector had the highest inhibitory efficiency , and com-pared with the nonsense group and control group , there was significant difference (all P<0.05).The expression level of ADAM17 mRNA and protein in the transfection group was significantly lower than that in the nonsense group and control group (all P<0.05).The cell proliferation activity and cell growth rate of the transfection group were significantly lower than those of the nonsense group and the control group (all P<0.05).The percentage of cells in S phase and G 2/M phase decreased in the transfection group , and most of the cells stayed in G0/G1 phase, which were significantly different from those in the control group and nonsense group (all P<0.05).Conclusion ADAM17-shRNA can inhibit the proliferation of human breast cancer MCF-7 cells, and the mechanism is related to its silencing effect on ADAM 17 gene.

Objective To investigate the effects of shRNA targeting a disintegrin and metalloproteinases 17(ADAM17)on the proliferation of human breast cancer MCF-7 cells in hypoxia environment.Methods The four specific ADAM17-shRNA se-quences aiming at ADAM17 were designed,transfected into MCF-7 cells by electroporation,and cultured in hypoxia environment. The experiment was divided into the control group (blank phosphate buffer solution,PBS),nonsense sequence group (transfected with ADAM17-shNC)and shRNA transfection group (transfected with ADAM17-shRNA,the highest silencing efficiency of shR-NA was selected for following experiments).Real-time PCR was used to detect the expression of ADAM17 mRNA.The prolifera-tion ability and cell cycle change of MCF-7 cells were detected by iCELLigence and flow cytometry (FCM),respectively.Results Compared with control group and nonsense sequence group,the four ADAM17-shRNA transfection groups all had the silence effect on ADAM17 gene expression (P <0.05 ),the difference was statistically significant(P <0.05 ),particularly shRNA1219 had the highest inhibitory rate (F =5.11,P <0.01 ).The cellular proliferation ability and cell growth speed in the shRNA transfection group were significantly decreased compared with the control group and nonsense sequence group (P <0.05).Most cells of shRNA transfection group remained in the G0/G1 phase (73.35 ± 2.45 ),which in the control group and nonsense sequence group was (62.56±2.35)and (62.68 ±1.20)respectively,the difference was statistically significant(P <0.05).The cell cycle progression was significantly delayed.Conclusion ADAM17-shRNA inhibits the proliferation of MCF-7 cells under hypoxic environment.

目的 利用shRNA干扰沉默人乳腺癌细胞MCF-7的解聚素-金属蛋白酶17(adisintegrin and metalloproteinases 17,ADAM17)基因,观察其对细胞增殖的影响.方法 针对ADAM17基因设计合成具有特异性的ADAM17-shRNA,经脂质体LipofectamineTM 2000转染MCF-7细胞.实验设对照组(空白PBS)、干扰组(转染干扰无义序列ADAM17-shNC)、实验组(转染ADAM17-shRNA),采用Reahime PCR检测各组细胞AD-AM17 mRNA的表达水平,四甲基偶氮唑盐比色法(MTT)检测细胞的增殖活性,流式细胞仪检测细胞周期变化.结果 实验组ADAM17 mRNA的相对表达量显著低于对照组和干扰组,差异有统计学意义(P＜0.05);实验组的MCF-7细胞增殖活性较其他两组明显降低,差异有统计学意义(P＜0.05);实验组的MCF-7细胞进入S期(23.60±1.09)％和G2/M期(6.30±0.82)％的比例降低,绝大多数细胞停留在G0/G1期(65.17±1.35)％,细胞周期延缓,与对照组、干扰组相比较差异具有显著性意义(P＜0.05).结论 ADAM17-shRNA对人乳腺癌细胞MCF-7的ADAM17基因具有沉默作用,从而抑制MCF-7细胞的增殖活性,延缓细胞周期进展.