CDKN3 knockdown reduces cell proliferation,invasion and promotes apoptosis in human ovarian cancer

Cyclin-dependent kinase inhibitor 3 (CDKN3) has been reported to promote tumor genesis. The aim of this study is to investigate the possible mechanisms of silence of CDKN3 exerting the suppressive role on epithelial ovarian cancer (EOC). To study the potential function of CDKN3 enrolled in the regulation of ovarian tumor, we monitored the EOC cells SKOV3 and HO8910 behaviors including proliferation, cell cycle, apoptosis and invasion. First, we found that CDKN3 was frequently over-expressed in EOC. Functional studies showed that silence of CDKN3 inhibited cancer cell proliferation by promoting cell cycle progression in G1 phase, decreased cell invasion and promoted EOC cells apoptosis. Western blot analysis of CDKN3-silence cells revealed down-regulation of DNA-replication and cell cycle related proteins. And, a significant correlation level of CDKN3 was observed which has been demonstrated to be a novel oncogene. These findings indicated that CDKN3 might serve as a useful potential target for treatment of ovarian cancer.     

MicroRNA-101 regulates the viability and invasion of cervical cancer cells

Background: Cervical cancer has the second highest morbidity and mortality rates of any malignancy in women worldwide, and it is one of the leading causes of death in Uygur women in Xinjiang China. MicroRNAs are involved in cancer development and progression. Previously, we found that miR-101 is significantly down-regulated in cervical cancer tissues from Uyghur women. The underlying pathophysiology and relevance to tumorigenesis of miR-101 is still largely unknown. The purpose of this study was to elucidate the molecular mechanisms of miR-101 regulation of cervical cancer cell viability and invasion. Materials and methods: The expression of miR-101 in cervical cancer cell line (SiHa) was detected by real-time PCR. A miR-101 mimic was overexpressed in SiHa cells, and MTT assays were performed to determine the impact on cell proliferation. Cell would heal assays and flow cytometry were used to detect migratory ability and cellular apoptosis, respectively. Immunohistochemistry was performed to assess protein expression of the miR-101 target gene COX-2. Results: MiR-101 was endogenously expressed in SiHa cells, and alterations in its expression had profound effects on cellular migration and invasion efficiency. Overexpression of miR-101 decreased proliferation in the MTT assay (the mimics at 490 nm absorbance is lower 60% than normal, and decreased cellular motility in the cell would healing assay (transfected: 37 ± 2 m, pre-transfected 184 ± 2 m). Apoptosis rate was significantly higher with overexpression of miR-101 relative to control (transfected: 76.6%, pre-transfected: 3.5%) (P < 0.05). The expression of Cox-2 was decreased in transfected cells. Conclusions: MiR-101 likely acts as a tumor suppressor in cervical cancer. Overexpression of miR-101 decreased expression of its target gene Cox-2 and inhibited proliferation and invasion, and promoted apoptosis to suppress tumorigenicity. MiR-101 is a promising new target for the development of therapeutic strategies for the clinical treatment of cervical cancer.     

小干扰RNA抑制RUNX2对人乳腺癌MCF-7细胞增殖、凋亡与侵袭的影响

目的探讨小干扰RNA抑制人RUNT相关转录因子2(RUNX2)对人乳腺癌MCF-7细胞增殖、凋亡与侵袭的影响。方法应用化学合成小干扰RNA技术沉默人乳腺癌MCF-7细胞中RUNX2基因的表达,用逆转录-聚合酶链反应(RT-PCR)和Western blot实验验证基因沉默效率。应用MTT方法检测细胞增殖能力变化,流式细胞仪检测细胞凋亡的变化,Transwell实验检测细胞侵袭能力变化。结果设计的siRNA能够明显抑制人乳腺癌MCF-7细胞RUNX2的表达。RUNX2基因沉默后,人乳腺癌MCF-7细胞的增殖能力降低,细胞凋亡率明显提高,侵袭能力显著降低(P<0.01)。结论RUNX2基因是治疗人乳腺癌MCF-7细胞的潜在靶点。     

糖蛋白非转移性黑色素瘤蛋白B对人肝癌细胞增殖、凋亡和侵袭能力的影响及其分子机制

 目的：研究糖蛋白非转移性黑色素瘤蛋白B（glycoprotein nonmetastatic melanoma protein B,GPNMB）对肝癌细胞增殖、凋亡和侵袭能力的影响及其分子机制。方法：以人HepG2细胞为研究对象,构建GPNMB的小干扰RNA（small interfering RNA,siRNA）以及过表达载体,转染入细胞后分别采用MTT法、流式细胞术和Transwell小室法观察其对HepG2细胞增殖、凋亡及侵袭能力的影响。结果：增殖实验结果表明GPNMB可促进HepG2细胞的增殖;流式细胞术检测发现GPNMB对HepG2细胞凋亡几乎无影响;细胞侵袭结果表明GPNMB可促进HepG2细胞的侵袭;当整合素β1亚基基因被siRNA沉默后,GPNMB对HepG2细胞增殖和侵袭的促进作用均受到明显抑制。结论: GPNMB可能通过与整合素β1亚基相互作用促进肝癌细胞的增殖和侵袭能力,提示GPNMB可以作为治疗肝癌的潜在靶点。     

PUMA增强依托泊苷诱导宫颈癌细胞凋亡

目的探讨PUMA蛋白在依托泊苷诱导的宫颈癌细胞凋亡中的作用。方法采用MTT方法检测依托泊苷对宫颈癌细胞系SiHa细胞活力的影响。通过Annexin-v／PI双染检测和光学显微镜形态观察检测依托泊苷对宫颈癌SiHa细胞凋亡的影响。通过转染siRNA干扰PUMA的表达,检测低表达PUMA后,宫颈癌细胞对依托泊苷的敏感性。通过免疫印迹方法检测PUMA干扰后PUMA的表达水平。结果依托泊苷显著抑制宫颈癌细胞的活力和诱导其凋亡,且随着浓度和时间的增加而细胞活力依次减少而凋亡率依次升高,且光学显微镜下有明显的细胞皱缩等凋亡形态。当转染PUMA的siRNA后．依托泊苷诱导的细胞凋亡显著减少。结论PUMA在依托泊苷诱导宫颈癌细胞凋亡中起了重要作用,它能够增强宫颈癌细胞对依托泊苷的敏感性。     

西格列汀抑制膀胱癌细胞增殖和诱导细胞凋亡

目的观察西格列汀对膀胱癌细胞增殖和凋亡的影响。方法西格列汀处理膀胱癌细胞T24及5637后,MTS检测细胞的增殖活性;Western blot检测细胞内组织蛋白酶B以及凋亡相关蛋白的表达变化。结果西格列汀显著抑制了膀胱癌细胞的增殖,经西格列汀处理后,T24及5637细胞内组织蛋白酶B的蛋白表达水平明显下降,凋亡相关蛋白PARP的剪切体表达水平升高。结论西格列汀抑制膀胱癌细胞增殖和诱导癌细胞凋亡,同时降低细胞内组织蛋白酶B的蛋白表达。     

BCL2L10 protein regulates apoptosis/proliferation through differential pathways in gastric cancer cells

The reason for and consequences of BCL2L10 down-regulation in gastric carcinoma are poorly understood. Our aim was to investigate the function of the protein BCL2L10 in gastric carcinoma. We investigated BCL2L10 expression using quantitative real-time PCR and immunoblotting. The methylation status of the BCL2L10 gene promoter was examined by bisulphite sequencing in fresh gastric normal and carcinoma tissues. We studied apoptosis and proliferation regulation in gastric cancer cell lines using flow cytometry, fluorescence staining, murine xenografting and immunoblotting. Pathway inhibitors were applied to confirm the major pathways involved in apoptosis or proliferation regulation. We observed significant correlations between lower BCL2L10 expression and CpG island hypermethylation of the BCL2L10 gene promoter in gastric carcinoma, apoptosis induced by over-expressed BCL2L10 through mitochondrial pathways, and proliferation accelerated by BCL2L10 siRNA via the PI3K-Akt signalling pathway in gastric cancer cell lines. The pro-apoptotic effect of BCL2L10 and growth promotion by BCL2L10 siRNA in gastric cancer cells suggest that it may be a tumour suppressor.     

MST1过表达抑制宫颈癌细胞系SiHa的增殖、迁移和侵袭

目的探讨哺乳动物不育系20样激酶1(MST1)对子宫颈癌SiHa细胞增殖、迁移和侵袭能力的影响。方法Western blot检测正常宫颈上皮细胞H8与宫颈癌细胞SiHa中MST1的表达;构建p J3H-HA-MST1质粒并转染SiHa细胞系,Western blot检测MST1、Ki-67和MMP9的蛋白表达;MTS、划痕实验和Transwell分别检测细胞增殖、迁移和侵袭能力。结果 SiHa细胞的MST1蛋白表达水平明显低于H8细胞(P0.05);SiHa细胞转染MST1质粒后,MST1蛋白表达明显升高,而Ki-67和MMP9蛋白表达水平显著下降(P0.05),SiHa细胞的增殖被明显抑制(P0.05),同时细胞的迁移和侵袭能力也显著降低(P0.01)。结论 MST1过表达可以抑制宫颈癌细胞SiHa的增殖、迁移和侵袭能力。     

SERPINA3 promotes endometrial cancer cells growth by regulating G2/M cell cycle checkpoint and apoptosis

Endometrial carcinoma (EC) is the most common gynecologic cancer worldwide and is one of the leading causes of death in women. Therefore, it is urgent to elucidate the pathological mechanisms of EC. SERPINA3 is a member of the serpin super-family of protease inhibitors. Its aberrant expression has been observed in various tumor cells. However, its clinical significance and biological function in endometrial cancer remains unknown. In the present study, we demonstrated that SERPINA3 expression was significantly up-regulated in EC samples and was closely correlated with lower differentiation, higher stage, positive lymph node or vascular thrombosis and negative estrogen receptor (ER), indicating a poor prognosis. We then demonstrated that SERPINA3 promoted EC cells proliferation by regulating G2/M checkpoint in cell cycle and inhibited cells apoptosis, and we further uncovered that the pro-proliferative effect of SERPINA3 on EC was likely ascribed to the activation of MAPK/ERK1/2 and PI3K/AKT signaling. The results of our study may provide insight into the application of SERPINA3 as a novel predictor of clinical outcomes and a potential therapeutic target of EC.     

miR-503 inhibits cell proliferation and induces apoptosis in colorectal cancer cells by targeting E2F3

Objective: Colorectal cancer (CRC) is one of the major healthcare problems worldwide. A lot of miRNAs are aberrantly expressed in CRC and involved in its development and progression. The purpose of this study was to investigate the expression and function of miR-503 in CRC. Methods: miR-503 expression was detected in CRC tissues and cell lines by Quantitative real-time PCR. Cell proliferation was assessed by MTT assay. Cell apoptosis and cell cycle distribution were measured by flow cytometry. Moreover, luciferase reporter assay and western blot were performed to determine the potential target of miR-503 in CRC cells. Results: miR-503 was significantly decreased in CRC tissues and cell lines in comparison with controls. Overexpression of miR-503 in CRC cells remarkably inhibited cell proliferation and induced apoptosis. Furthermore, E2F3 was identified as a direct target of miR-503 in CRC cells and down-regulation of E2F3 had a similar effect as miR-503 overexpression on CRC cells. In addition, the expression of E2F3 was negatively correlated with miR-503 level in CRC tissues. Conclusions: miR-503 inhibits cell proliferation and induces apoptosis by directly targeting E2F3 in CRC cells, indicating its potential application in CRC diagnosis and therapy.     

Effect of bone morphogenetic protein-2 on proliferation and apoptosis of gastric cancer cells

Objective: To investigate the effects of bone morphogenetic protein-2 (BMP-2) on the proliferation, differentiation and apoptosis of normal human gastric mucosal cells and gastric cancer cells.Methods: Poorly differentiated gastric cancer BGC823 cells, moderately differentiated gastric cancer cells and normal human gastric mucosal epithelial GES-1 cells were independently treated with recombinant human BMP-2 or its inhibitor Noggin. MTT assay was performed to detect the proliferation, flow cytometry done to measure the cell cycle and apoptosis and immunohistochemistry carried out to determine the expression of cyclin-dependent kinase 4 (CDK4).Results: BMP-2 exerted inhibitory effect on the growth of all types of cells and the inhibition become more evident with the increase of BMP-2 dose. After treatment with 200 ng/ml BMP-2, cancer cells arrested in G1 phase and those in S phase reduced. Gastric cancer cells had higher CDK4 expression than GES-1 cells. BMP-2 decreased CDK-4 expression in cancer cells but had no influence in GES-1 cells. Noggin conferred promotive effect on the growth of 3 types of cells. In 2 types of cancer cells, treatment with 2000 ng/ml Noggin significantly increased the proportion of cells in S phase but reduced that in G1 phase. However, Noggin did not affect the cell cycle of GES-1 cells. The CDK4 expression was markedly increased in 2 types of cancer cells but that of GES-1 remained unchanged after treatment with 2000 ng/ml Noggin.Conclusions: BMP-2 may inhibit the proliferation of both normal and malignant gastric epithelial cells, down-regulate CDK4 expression in gastric cancer cells and arrest gastric cancer cells in G1-phase in cell cycle. Through antagonizing BMP-2, Noggin, may accelerate the proliferation of gastric cancer cells. Thus, the abnormality of BMP signaling pathway may play an important role in the pathogenesis of gastric cancer.     

PXMP4促进结直肠癌细胞增殖、迁移和侵袭

目的 探讨过氧化物酶体膜蛋白4(peroxisomal membrane protein 4,PXMP4)对结直肠癌细胞增殖、迁移和侵袭能力的影响.方法 采用生物信息学分析65例配对结直肠癌组织中PXMP4 mRNA的表达.应用免疫组化检测112例配对结直肠癌组织中PXMP4的表达,并进行临床病理相关分析;利用RT-P...     

白藜芦醇诱导人宫颈癌细胞凋亡的作用

目的观察白藜芦醇对人宫颈癌Hela细胞凋亡的影响。方法采用TUNEL法检测不同浓度的白藜芦醇诱导体外培养的Hela细胞凋亡情况。结果浓度大于50μmol／L的白藜芦醇处理Hela细胞后,细胞凋亡特征明显可见,并呈时间和剂量依赖关系。结论白藜芦醇对人宫颈癌Hela细胞有明显的促凋亡作用。     

Tumor‐associated lymphatic endothelial cell promotes invasion of cervical cancer cells

The most common way for cervical cancer to spread is through the lymphatic system. Tumor‐associated lymphatic endothelial cell (TLEC) has been considered to play a crucial role in metastasis of certain cancers. The aim of this study was to isolate TLEC from human cervical cancers and explore its involvement in metastasis‐associated behaviors in vitro. Lymphatic vessels in 62 cervix tissue specimens ranging from cervical intraepithelial neoplasia (CIN) to advanced invasive cancer were detected using immunochemical staining with D2‐40 antibody. Relation of lymphatic vessel density (LVD) to clinicopathological characters was analyzed. Primary TLECs were isolated by LYVE‐1 immuno‐magnetic beads from cervical cancer tissues and verified through expression of LEC markers Prox‐1 and D2‐40, and then cultured in vitro. Invasiveness and viability of cells were assessed by transwell assay and typan blue exclusion, respectively. Our results showed that higher LVD was significantly associated with advanced FIGO stage, pelvic lymphatic nodal metastasis (LNM), and poorer cell differentiation. TLECs were successfully primarily isolated and cultured in vitro. Supernatant of TLEC enhanced invasiveness of Hela cell, but did not significantly affect cell viability. In conclusion, TLECs might actively promote lymphatic metastasis of cervical cancer. Further studies are needed to demonstrate the underlying mechanisms.     

miR-101 suppresses tumor proliferation and migration,and induces apoptosis by targeting EZH2 in esophageal cancer cells

Aim: To investigate the role of miR-101 in the regulation of tumor proliferation, invasion, apoptosis and to its target gene in human ESCC. Methods: The expression level of miR-101 in Eca109 cell line was determined by real-time polymerase chain reaction (PCR). After transfected with miR-101 mimics and inhibitor, proliferation, migration and apoptosis in ESCC cell line (Eca109) were detected by MTT, cell wound healing assay and flow cytometry, respectively. The expression of EZH2 in Eca109 cell was examined by immunohistochemical staining. Results: We found that miR-101 was significantly down-regulated in ESCC cell than in matched normal esophageal epithelium cell. The expression level of miR-101 was inversely correlated to EZH2 protein expression in ESCC cell. In Eca109 cells, over-expression of miR-101 significantly inhibited the migration and invasion of ESCC cells, and promotes cell apoptosis. Conclusions: These findings suggest that decreased expression of miR-101 might promote metastasis of human ESCC by inducing accumulation of EZH2 protein.     

DDX3表达上调在人宫颈癌细胞增殖中的作用分析

目的:探讨DDX3表达上调在人宫颈癌细胞增殖中的作用。方法:采用免疫组织化学方法检测2012年4月至2013年3月河南省人民医院59例宫颈癌组织及癌旁非肿瘤组织标本中的DDX3表达情况。同时,以宫颈癌细胞HeLa为研究对象,通过慢病毒介导的宫颈癌细胞过度表达来检测DDX3的功能。采用细胞增殖与活性检测试剂盒评估细胞存活...     

膜联蛋白A2对人宫颈癌HeLa细胞增殖、迁移和凋亡的影响

 目的：研究膜联蛋白A2（annexin A2, ANXA2）对人宫颈癌HeLa细胞增殖、迁移和凋亡能力的影响。方法：以HeLa细胞为研究对象,构建过表达载体以及ANXA2-siRNA,转染入细胞。将细胞分为正常对照组、scrambled组、ANXA2过表达组及ANXA2-siRNA组。应用real-time PCR法检测ANXA2 mRNA表达水平及Western blotting检测ANXA2蛋白表达水平。分别采用MTT法、Boyden小室法和流式细胞术观察ANXA2对HeLa细胞增殖、迁移及凋亡能力的影响。结果：ANXA2过表达组可以显著促进HeLa细胞的增殖和迁移;ANXA2-siRNA组明显抑制HeLa细胞的增殖和迁移;ANXA2对HeLa细胞凋亡几乎无影响。结论:沉默ANXA2对人宫颈癌细胞的凋亡无显著影响,但可显著抑制其增殖能力和迁移能力。ANXA2可能在宫颈癌的发生发展中具有十分重要的作用,提示它有可能成为宫颈癌治疗的分子靶点。     

康莱特注射液对胃癌患者细胞凋亡、增殖及T细胞亚群的影响

目的：探讨康莱特对胃癌细胞凋亡对增殖及胃癌患者T细胞亚群的影响。方法：进展期胃癌患者30例，随机分为3组（每组各10例）：A组（对照组）：手术前后常规给予全胃肠外营养（total parenteral nutrition,TPN)治疗；B组（康莱特治疗组）：术前5d及术后9d，给予康莱特注射液200mL/d静脉滴入加常规TPN治疗；C组（化疗组）：术前给予5d化疗，甲酰四氢叶酸钙（calcium folinatefor,CF)200mg及5氟脲嘧啶（5－FU）750mg/d静脉滴注加常规TPN治疗。分别于治疗前及术后1d,5d及10d，采集外周静脉血，应用免疫荧光法检测CD3^ 、CD4^ 、CD8^ T细胞亚群。手术中取胃癌病理组织，用末端转移酶介导的dUTP切口末端标记法和免疫组织化学染色法检测胃癌细胞的凋亡（AI）与增殖（PI）及二者之比（AI／PI）。结果：B组与C组相比较，胃癌细胞的AI、PI及AI／PI，无显著差异（P＞0．05）；B组与A组相比较，上 3种指标则具有显著差异（P＜0．01）。治疗前3组CD3^ 、CD4^ 、CD8^ T细胞亚群的百分率无显著差异（P＞0．05）；术后1d,5d3组CD3^ 、CD4^ 、CD8^ T细胞亚群的百分率差异显著（P＜0．01）；术后10d,CD3^ 、CD4^ 、CD8^ T细胞亚群的百分率，B组与A组以及B组与C组相比较，差异性分别为显著（P＜0．05）及非常显著（P＜0．01）。结论：康莱特可明显促进胃癌细胞凋亡和抑制其增殖，有助于提高围手术期患者的免疫功能。