TROP-1/Ep-CAM and CD24 are potential candidates for ovarian cancer therapy

To clarify the possible roles of epithelial cell adhesion molecule (TROP-1/Ep-CAM) and CD24 molecule (CD24) in ovarian tumorigenesis, and explore the possible mechanism underlying this disease. Recombinant eukaryotic expression vectors pCIneo-TROP-1/Ep-CAM and pCIneo-CD24 were transfected into human normal ovarian surface epithelia cell line IOSE-80 respectively, with IOSE-80 cells transfected with the empty vector pCIneo as control. MRNA and protein expression of TROP-1/Ep-CAM and CD24 were detected by RT-PCR and Western blotting, respectively. Cell migration was assayed by trans-well inserts; cell proliferation and adhesion were analyzed by CCK-8 Cell Counting kit; cell cycle and cell apoptosis analysis were performed by flow cytometer. The expressions of TROP-1/Ep-CAM and CD24 were obviously up-regulated in TROP-1/Ep-CAM group and CD24 group compared to that in control group (P < 0.01). Cells of TROP-1/Ep-CAM group and CD24 group was significantly promoted migratory and proliferation abilities, but inhibited cell apoptosis and adhesive than that of control group (P < 0.05). Besides, the number of the cells in G1 and G2 stages was significantly lower in two disease groups than that in control group (P < 0.05). TROP-1/Ep-CAM and CD24 may play key roles in the progression of ovarian cancer through promoting migration, proliferation, inhibiting cell apoptosis and adhesion, and disturbing cell cycle. They may be used as specific therapeutic targets in the treatment of ovarian cancer. However, further experiments are still needed to confirm our results.     

Mechanism underlying the regulation of lncRNA ACTA2-AS1 on CXCL2 by absorbing miRNA-532-5p as ceRNA in the development of ovarian cancer

Objective: To explore the mechanism underlying the regulation of long non-coding RNA (LncRNA) ACTA2-AS1 on CXCL2 as a ceRNA of miR-532-5p in the progression of ovarian cancer (OC). Methods: A qRT-PCR assay was carried out for analyzing the expression changes of ACTA2-AS1, miR-532-5p, as well as CXCL2 in OC tissues and corresponding healthy paracancerous tissues HOSEpiC (human ovarian epithelial cells), and OC cells. OC cells were grouped and transfected, and the fluorescent in situ hybridization was adopted for evaluating ACTA2-AS1 in the cells. Additionally, a dual luciferase reporter (DLR) assay was carried out for verifying the correlation of ACTA2-AS1 with miR-532-5p and of miR-532-5p with CXCL2. Cells were transfected with si-ACTA2-AS1, miR-532-5p, or CXCL2 overexpression plasmids, and then the cell proliferation, invasion, and apoptosis were determined using MTT, Transwell, and flow cytometry assays, respectively. Results: Compared with paracancerous tissues and HOSEpiC cells, OC tissues and cells showed increased ACTA2-AS1 and CXCL2 expression and decreased miR-532-5p expression (all P<0.05). ACTA2-AS1 acted as ceRNA in OC by negatively regulating miR-532-5p. Additionally, upregulating ACTA2-AS1 intensified the proliferation and invasion of cancer cells and suppressed their apoptosis (all P<0.05), and inhibition of it resulted in opposite results. In contrast, overexpressing miR-532-5p suppressed the proliferation, invasion, and clone formation of the cells and promoted their apoptosis (all P<0.05). The effect of ACTA2-AS1 on OC cells can be partially reversed by overexpressing miR-532-5p. Moreover, CXCL2, positively correlated with ACTA2-AS1 in expression (P<0.0001, r=0.7385), was the target of miR-532-5p, and its overexpression could partially offset the influence of miR-532-5p on OC cells. Conclusion: LncRNA ACTA2-AS1 can act as a tumor promoter in OC by absorbing miR-532-5p as ceRNA and regulating CXCL2, and ACTA2-AS1 inhibitor is expected to play a role in targeted therapy of OC.     

Diagnostic significance of miR-106a in gastric cancer

As one of the most common malignant tumors, gastric cancer still lacks tumor markers with enough specificity and sensitivity. Therefore the development of novel tumor markers is necessary for early diagnosis in clinics. MicroRNA (miR) has been known to be of unique expressional patterns in various tumors and may work as potential tumor markers for clinical use. This study thus explored the significance of plasma miR-106a in clinical diagnosis of gastric cancer and its effects on proliferation of cancer cells. Plasma miR-106a levels were quantified by real-time quantitative fluorescent PCR methods in 80 cases of gastric cancer patients and healthy individuals to analyze the correlation between miR-106a and clinical features. MiR-106 inhibitor was further transfected into human gastric carcinoma cells for further cell proliferation using CCK-8 approach. MiR-106a was significantly up-regulated in gastric cancer patient plasma samples compared to healthy individuals (P<0.01). The area under ROC curve was 0.895 (95% CI: 0.846~0.943). It has a specificity of 93.8% and a sensitivity of 77.5% in diagnosing gastric cancer. MiR-106a level was also correlated with cancer differentiation stage, lymph node metastasis, TNM stage and tumor size (P<0.05). The down-regulation of miR-106 in gastric carcinoma cells inhibited cell proliferation (P<0.05). MiR-106a was significantly up-regulated in gastric cancer patients and can facilitate the in vitro proliferation of tumor cells. It may work as a biological marker for gastric cancer.     

Cyclic RNA Circ_0000735 sponges miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis

Objective: The objective of this study was to investigate the effect on the proliferation, invasion, and apoptosis of bladder cancer cells through miR-502-5p of the Circ_0000735 circular RNA. Methods: Circ_0000735 and miR-502-5p expression of bladder cancer patients in malignant and paracancerous tissues was identified using qRT-PCR. Nucleoplasm isolation assay and RNase R enzymatic assay were used to classify Circ_0000735 subcellular origin and stability. Dual luciferase reporter assay and RIP assay were used to confirm Circ_0000735 and miR-502-5p targeting relationships. Cell proliferation, apoptosis, and invasion capacity were identified using CCK8, flow cytometry, and transwell assays. To confirm the effect of Circ_0000735 on tumorigenesis in nude mice, in vivo experiments were conducted. Results: Circ_0000735 expression was increased in bladder cancer tissues and cells compared with paraneoplastic tissues and normal cells, and miR-502-5p expression was reduced (both P<0.05). In the cytoplasm, Circ_0000735 was largely clustered and could not be digested by the RNase R enzyme, and ceRNA may play a role in bladder cancer cells. Circ_0000735 silencing prevented cell proliferation and invasion and facilitated apoptosis (all P<0.05). The incorporation of miR-502-5p inhibitor rescued the effect on bladder cancer cells of Circ_0000735 silencing. In vitro experiments showed that inhibition of Circ_0000735 expression was beneficial in suppressing tumorigenic ability in nude mice. Conclusion: Circ_0000735 can adsorb miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis. Circ_0000735 may be an effective molecular target for bladder cancer therapy.     

Long non-coding RNA TP73-AS1 predicts poor prognosis and regulates cell proliferation and migration in cervical cancer

IntroductionCervical cancer is one of the most common malignant tumors in women, which seriously affects women’s health, especially in developing countries. This study aims to investigate novel molecular markers for poor prognosis of cervical cancer to achieve correct guidance of clinical treatment, accurate assessment of prognosis, and provide a new basis for the choice of reasonable treatment options for cervical cancer patients.Material and methodsQRT-PCR was employed to investigate the expression of lncRNA TP73-AS1 in cervical cancer tissues and cell lines. COX multivariate analysis showed the relationship between TP73-AS1 expression and clinicopathological features of patients with cervical cancer. Colony formation and MTT assay detected the effect of TP73-AS1 on proliferation of cervical cancer cells. The effect of TP73-AS1 on migration and invasion of cervical cancer cells was determined by the wound-healing assay and transwell assay. Western blot was performed to assess the expression of EMT markers.ResultsThis study showed that lncRNA TP73-AS1 was up-regulated in cervical cancer tissues and cell lines (p < 0.001), and high expression of TP73-AS1 could be considered as an independent prognostic factor (p < 0.05). Moreover, lncRNA TP73-AS1 promotes cervical cancer cell migration and invasion, and knockdown of TP73-AS1 inhibits the growth of cervical cancer cells (p < 0.001).ConclusionsOur results indicated that lncRNA TP73-AS1 was up-regulated in cervical cancer tissues and cell lines, predicting poor prognosis of cervical cancer and regulating cell proliferation and migration.     

JMJD5 is a potential oncogene for colon carcinogenesis

Objective: To observe the effects of Jumonji C domain-containing (JMJD) 5 depletion on colon cancer (CC). Methods: A short-hairpin RNA targeting JMJD5 was transfected into a lentivirus to make Lv-shJMJD5 for infection into the Caco-2 human cell. Besides, a negative control shRNA was constructed. The mRNA and protein levels of JMJD5 were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting, respectively. Cell proliferation, migration, and invasion were assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), soft agar colony assay and transwell assay, respectively. In addition, immunohistochemical (IHC) staining was performed to investigate the expression of JMJD5 in adjacent normal tissues and tumor tissues from patients with CC. Results: Compared with control group, mRNA and protein levels of JMJD5 was significantly reduced after infection with Lv-shJMJD5 (P<0.05), and Caco-2 cell proliferation, migration, and invasion were all obviously inhibited (P<0.05). The results of IHC showed that JMJD5 was significantly up-regulated compared with normal tissues (P<0.01). Additionally, follow-up data demonstrated that the survival rate of patients with high expression of JMJD5 was obviously lower than that with low expression (P<0.01). Conclusions: JMJD5 depletion could significantly inhibit human CC cell proliferation, migration, and invasion, implying that JMJD5 might be a potential oncogene.     

Adsorption of miR-218 by lncRNA HOTAIR regulates PDE7A and affects glioma cell proliferation,invasion, and apoptosis

Objective: To evaluate the role of targeted adsorption of miR-218 by long-chain non-coding RNAHOTAIR to regulate PDE7A on glioma cell proliferation, invasion, and apoptosis. Methods: The expressions of lncRNA HOTAIR, miR-218, and PDE7A in glioma tissues and normal parcancer tissues, NHA and glioma cell lines were determined, and correlations among the three genes were analyzed. The subcellular localization of lncRNA HOTAIR was determined by fluorescent in situ hybridization. Dual-luciferase reporter assay was used to validate the targeted relationship between lncRNA HOTAIR/miR-218/PDE7A. Glioma cells were grouped to receive intervention of lncRNA HOTAIR or miR-218. MTT, transwell, and flow cytometry were performed to determine the proliferation, invasion, and apoptosis of cells. Results: Compared with the normal tissues and cells, the expression of lncRNA HOTAIR was increased while miR-218 was suppressed in glioma tissues samples and cells (all P<0.05). Inhibition of lncRNA HOTAIR expression, was able to induce apoptosis and suppress the proliferation and invasion of cells (all P<0.05). LncRNA HOTAIR is mainly localized in the cytoplasm, and is able to adsorb miR-218 as ceRNA. The effect of knockdown of HOTAIR on glioma cells could be partially rescued by miR-218 inhibitor. The expression of PDE7A was enhanced in glioma tissues and cells compared to normal tissues and cells (all P<0.05), which positively correlated with the expression of HOTAIR (r=0.546, P<0.05) and negatively correlated with the expression of miR-218 (r=0.363, P<0.05). The targeted relationship between miR-218 and PDE7A was validated: Overexpression of miR-218 was able to suppress the proliferation and invasion of glioma cells and restrain apoptosis compared to the miR-NC group (all P<0.05). The effect of miR-218 on glioma cells could be partially rescued by PDE7A. Conclusion: lncRNA HOTAIR can adsorb miR-218 to regulate expression of PDE7A and promote the malignant biologic behavior of glioma cells.     

Potential role of CXCR3 in proliferation and invasion of prostate cancer cells

Aim: To investigate the potential role of CXCR3 expression on prostate cancer cell proliferation and invasion and to illustrate its mechanism. Methods: Human PC-3 cells were transfected with siRNA-CXCR3A and siRNA-CXCR3B plasmids respectively. The mRNA expressions of CXCR3A and CXCR3B in PC-3 cells from each group were analyzed using RT-PCR. Besides, cell proliferation ability and cell invasion ability of PC-3 cells in each group were analyzed using MTT assay and Matrige assay respectively. Additionally, expressions of CXCR3 downstream proteins were detected using Western blotting. Results: mRNA level of CXCR3A was decreased while CXCR3B mRNA level was increased in PC-3 cells (P<0.05). Compared with the controls, down-regulation of CXCR3A but up-regulation of CXCR3B significantly inhibited PC-3 cell proliferation and cell invasion ability (P<0.05). Besides, aberrant CXCR3 expression significantly increased expressions of phospholipase C (PLCβ), matrix metallo proteinase (MMP-1), and MMP-3 except MMP-7 in PC-3 cells (P<0.05). Conclusion: The data presented in our study suggested that aberrant CXCR3 expression may play crucial roles in suppressing PC metastasis via inhibiting cell proliferation and invasion ability through the PCLβ signaling pathway.     

UCH-LI acts as a novel prognostic biomarker in gastric cardiac adenocarcinoma

Gastric cardiac adenocarcinoma (GCA) accounts for a majority of gastric cancer population and harbors unfavorable outcome. Ubiquitin C-terminal hydrolase L1 (UCH-L1) belongs to the deubiquitinating enzyme family, which could regulate cell growth in human cancers. In the present study, expression of UCH-L1 was evaluated in 196 GCAs by immunohistochemistry using tissue microarray and its function on gastric cancer cells was measured. UCH-L1 expression was increased in GCA specimens, compared with their normal tissues and UCH-L1 overexpression is tightly correlated with tumor size and overall TNM stage. Log-rank analysis showed that UCH-L1 positive is reversely associated with cumulative survival (P<0.001). Multivariate Cox regression model showed that UCH-L1 overexpression is a remarkably negative predictor in GCA prognosis (Hazard Ratio=0.53, P<0.01), along with advanced TNM stage that is a known negative factor in gastric cancers (Hazard Ratio=0.33, P<0.05). Silencing of UCH-L1 reduced the ability of cell proliferation, colony formation, migration and invasion of gastric cancer cells. Our findings suggest that UCH-L1 is a promising prognostic biomarker for GCAs and might play an important role in the carcinogenesis of gastric cancer.     

布托啡诺调控PBX3基因抑制乳腺癌细胞系MCF7增殖、迁移和侵袭

目的探讨布托啡诺(butorphanol)对乳腺癌细胞增殖、迁移和侵袭的影响以及相关的分子机制。方法用MTT法检测不同浓度布托啡诺对人乳腺癌细胞系MCF7的抑制作用;用Transwell迁移及侵袭实验检测不同浓度布托啡诺对人乳腺癌MCF7细胞迁移及侵袭的影响;RT-qCR与Western blot法分别检测乳腺癌细胞系、正常乳腺上皮细胞以及布托啡诺对MCF7细胞中PBX3 mRNA及蛋白表达的影响;观察转染si-PBX3或si-con后,MCF7细胞增殖、迁移及侵袭能力的变化;PBX3过表达验证布托啡诺对乳腺癌增殖、迁移及侵袭的作用机制。Western blot检测cyclin D1和MMP-2蛋白表达。结果PBX3在乳腺癌细胞系中的表达上调,沉默PBX3表达可明显抑制MCF7细胞增殖、迁移及侵袭,同时抑制cyclin D1和MMP-2的表达;不同浓度的布托啡诺干预能显著抑制MCF7细胞增殖、迁移及侵袭且具有浓度依赖性,还可抑制PBX3、cyclin D1和MMP-2的表达;过表达PBX3可逆转布托啡诺对乳腺癌细胞增殖、迁移及侵袭的抑制作用。结论布托啡诺可通过抑制PBX3降低乳腺癌细胞增殖、迁移及侵袭能力。     

长链非编码RNA PVT1调控miR-551通过Wnt信号通路对卵巢癌迁移和侵袭的影响

目的:探讨长链非编码RNA PVT1在卵巢癌组织中的表达情况及其在卵巢癌细胞迁移和侵袭过程中的作用及机制。方法:q PCR检测卵巢癌和正常卵巢组织及不同卵巢癌细胞中PVT1的表达情况;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后卵巢癌细胞侵袭和迁移能力的变化;双萤光素酶报告基因检测PVT1与微小RNA(miR)-551的相互作用;Transwell侵袭实验和细胞划痕实验分别检测沉默PVT1后miR-551-inhibitor对卵巢癌细胞侵袭和迁移能力的影响;Western blot法检测沉默PVT1后Wnt信号通路相关蛋白的表达情况。裸鼠皮下成瘤实验检测沉默PVT1对卵巢癌成瘤重量及体积的影响。结果:与正常卵巢组织相比,卵巢瘤组织中PVT1表达明显增高(P0.05);卵巢癌细胞株ES-2中PVT1表达水平最高(P0.05);沉默PVT1可以抑制卵巢癌细胞侵袭和迁移能力;PVT1能与miR-551的位点特异性结合;沉默PVT1后,miR-551-inhibitor可以促进卵巢癌细胞侵袭和迁移能力;沉默PVT1后Wnt信号通路蛋白的表达相应下调;与阴性对照组相比,PVT1-siRNA组荷瘤小鼠肿瘤体积和重量都明显减小(P0.05)。结论:PVT1在卵巢癌发生发展过程中起重要作用,它可以靶向调节miR-551,通过Wnt信号通路调控卵巢癌细胞的侵袭和迁移能力。     

Desmocollin 3 mediates follicle stimulating hormone-induced ovarian epithelial cancer cell proliferation by activating the EGFR/Akt signaling pathway

Follicle-stimulating hormone (FSH) is associated with the pathogenesis of ovarian cancer. We sought to explore whether desmocollin 3 (Dsc3) mediates FSH-induced ovarian epithelial cancer cell proliferation and whether the EGFR/Akt signaling pathway may be involved in this process. Dsc3 positivity in ovarian tissue specimens from 72 patients was assessed by immunohistochemistry. The positive expression rates of Dsc3 were similar in ovarian cancer tissues (24/31:77.4%) and borderline ovarian tumor tissues (18/22:81.8%) (P>0.05), but were significantly higher in these cancerous tissues than in benign ovarian cyst tissues (3/19:15.8%) (P<0.05). Consistently, the expression of Dsc3 in four out of five ovarian cancer cells (HO8910, Skov3ip, Skov and Hey cells, but not ES-2 and in borderline ovarian MCV152 tumor cells was higher than in the immortalized ovarian epithelial cell line, Moody. FSH up-regulated the expression of Dsc3 and EGFR in a dose- and time-dependent manner. Furthermore, a converse relationship between the expression of Dsc3, EFGR and PI3K/Akt signaling was elucidated using RNA interference and PI3K/Akt inhibitor in the absence and presence of FSH. A role for these proteins in FSH-induced cell proliferation was verified, highlighting their interdependence in mediating ovarian cancer cell function. These results suggest that Dsc3 can mediate FSH-induced ovarian cancer cell proliferation by activating the EGFR/Akt signaling pathway.     

Knock-down of ABCE1 gene induces G1/S arrest in human oral cancer cells

Purpose: This study aims to explore the clinical characteristics of ATP binding cassette E1 (ABCE1) in oral squamous cell carcinomas (OSCC) and its roles in the proliferation, invasiveness, migration and apoptosis of the human oral squamous cell carcinoma cells CAL-27. Methods: The expression of ABCE1 and its target protein-RNase L, were first studied in tumor tissues of OSCC and adjacent non-tumor tissues. Moreover, CAL-27cells were transfected by ABCE1-specific shRNA, then MTT assay, the transwell and scratch assay were used to study cell proliferation and migration activity; the apoptosis rate and cell cycle distribution were tested by flow cytometry. Western blot and RT-PCR assay were adopted to measure their silencing efficacy. Results: ABCE1 expression is low in the adjacent non-tumor tissues while the expression is high in the oral cancer; the expression is reversely proportional to the differentiation degrees. The expression of RNaseL was in contrary to ABCE1. After transfected with ABCE1-siRNA, the proliferation, invasiveness and migration capabilities of cells decreased significantly whilst the apoptosis rate enhanced greatly (P < 0.01). Meanwhile, the expression of ABCE1 in CAL-27 cells was blocked (P < 0.01) while the expression of RNase L increased significantly (P < 0.01). Conclusion: ABCE1 is closely connected with the pathogenesis and development of oral cancer, which acts through the cellular pathways of 2-5A/RNase L.     

SFRP5基因沉默促进人类胰腺癌细胞系PANC-1增殖与迁移

目的探讨慢病毒介导短发夹RNA(shRNA)沉默SFRP5基因对人类胰腺癌细胞系PANC-1细胞增殖、侵袭和迁移能力的影响。方法构建靶向SFRP5基因特异性shRNA慢病毒载体并转染人胰腺癌PANC-1细胞系,以空白质粒转染阴性对照组,未处理细胞做为空白对照组。用real-time PCR及Western blot检测转染前后SFRP5 RNA以及蛋白的表达;CCK-8实验检测细胞体外增殖能力;使用Transwell小室实验分析细胞侵袭能力;细胞划痕实验分析细胞迁移能力。结果成功建立稳定转染shRNA-SFRP5胰腺癌PANC-1细胞株。SFRP5病毒转染组与阴性对照组及空白对照组相比细胞的增殖能力明显增加(P0.01);SFRP5病毒转染组的细胞侵袭、迁移能力明显高于阴性对照组及空白对照组(P0.01)。结论 SFRP5慢病毒干扰载体能有效抑制SFRP5基因在人胰腺癌PANC-1细胞中的表达,进而促进细胞的增殖、侵袭和迁移。     

Effect of apolipoprotein B mRNA-editing catalytic polypeptide-like protein-3G in cervical cancer

Cervical cancer is one of the most common gynecologic cancers. The role of apolipoprotein B mRNA-editing catalytic polypeptide-like protein-3G (APCBEC-3G) in cervical cancer has yet to be elucidated. This study intends to explore the effect ofAPCBEC-3G on cervical cancer cell proliferation and invasion. In vitro, the cervical cancer cell line Hela was transfected by APCBEC-3G plasmid. The mRNA and protein expression levels of APCBEC-3G were detected by Real-time PCR and Western blot, respectively. Cervical cancer cell proliferation was determined by MTT. Transwell assay was applied to measure the effect of APCBEC-3G on cell invasion. APCBEC-3G mRNA and protein increased significantly after transfection (P<0.05) and cervical cancer cell proliferation and invasive ability were decreased significantly (P<0.05). APOBEC-3G serves as a suppressor of cervical cancer cell proliferation and invasion. Our research provides theoretical basis for further investigationAPOBEC-3G effect in cervical cancer occurrence and development.     

Role of TM4SF1 in regulating breast cancer cell migration and apoptosis through PI3K/AKT/mTOR pathway

Purpose: The purpose of this study was to investigate the effect of transmembrane-4-l-six-family-1 (TM4SF1) on breast cancer cell line MDA-MB-231 invasion and apoptosis and its mechanism through PI3K/AKT/mTOR pathway. Methods: siRNA-TM4SF1 and pcDNA-TM4SF1 plasma were constructed and then transfected into MDA-MB-231 cells respectively. Real time (RT)-PCR was used to measure the mRNA expression of TM4SF1 in each group. Also, matrigel method and Annexin V-FITC were used to detect the effect of TM4SF1 expression on MDA-MB-231 cell migration and apoptosis respectively. Besides, western blotting analyze was used to assay the effects of TM4SF1 expression on PI3K/AKT/mTOR pathway associated proteins expressions. Results: The results showed that after being transfected with siRNA-TM4SF1, TM4SF1 expression was significantly declined, while it was significantly increased after cells were transfected with pcDNA-TM4SF1 (P<0.05). Compared with the controls, TM4SF1 overexpression significantly contributed MDA-MB-231 cell migration but decreased apoptotic cells (P<0.05), which were opposite to the results when TM4SF1 was sliced in cells. Moreover, TM4SF1 slicing significantly decreased the expressions of phosphorylated (p)-AKT, p-mTOR, and p-P70 (P<0.05). Conclusion: Our study suggested that TM4SF1 may be a therapeutic target for breast cancer treatment and may loan insight into the mechanisms behind the development and metastasis of advanced breast cancer.     

乙酰辅酶A羧化酶1对胶质瘤细胞系U87细胞增殖、迁移和侵袭的影响

目的 探讨乙酰辅酶A羧化酶1（ACC1）对人胶质瘤细胞系U87细胞增殖、迁移及侵袭的作用。 方法 Western blotting检测人胶质瘤细胞系U87、U251及U373中ACC1的表达;构建ACC1过表达质粒载体,将过表达ACC1质粒载体瞬时转染至U87细胞中;Western blotting检测转染后U87细胞中ACC1表达情况;MTT实验检测过表达ACC1对U87细胞增殖的影响;Transwell迁移和侵袭实验分别检测过表达ACC1对U87细胞迁移和侵袭的影响;划痕实验检测过表达ACC1对U87细胞划痕愈合能力的影响;Western blotting检测相关蛋白表达变化。 结果 与人胶质瘤细胞系U251和U373相比,U87细胞中ACC1表达较低;ACC1过表达抑制U87细胞增殖（P<0.01）;ACC1过表达抑制U87细胞迁移、侵袭和划痕愈合能力（P<0.01）;ACC1过表达迁移和侵袭相关蛋白波形蛋白(vimentin)、纤维连接蛋白(fibronectin)和尿激酶型纤溶酶原激活剂(uPA)表达下调（P<0.01）,凋亡抑制蛋白Bcl-2和细胞周期蛋白(cyclin) B、cyclin D表达下调（P<0.01）,p-STAT3蛋白表达下调（P<0.01）,细胞周期蛋白P21表达上调（P<0.01）。 结论 过表达ACC1可能通过抑制STAT3活性,抑制人胶质瘤细胞的增殖、迁移和侵袭。     

MiR-634 通过mTOR 通路抑制宫颈癌细胞c4-1、caski 增殖并促进其凋亡的研究

目的:探讨MiR-634 通过mTOR 通路抑制宫颈癌细胞c4-1、caski 增殖并促进其凋亡的相关机制。方法:选取30 例宫颈癌组织标本和宫颈癌细胞c4-1、caski 作为研究对象,分析正常组织和宫颈癌组织的MiR-634 和mTOR 表达,MiR-634对宫颈癌细胞mTOR 的表达水平以及对宫颈癌细胞的增殖、迁移和侵袭的影响。结果:宫颈癌组织MiR-634 相对表达水平显著低于正常组织(P<0.05),宫颈癌组织mTOR 相对表达水平显著高于正常组织(P<0.05),宫颈癌组织mTOR 的IS 得分显著高于正常组织(P<0.05);过表达MiR-634 显著降低mTOR 的RNA 和蛋白表达水平(P<0.05),抑制MiR-634 的表达显著提高mTOR 的RNA 和蛋白表达水平(P<0.05);过表达MiR-634 显著降低宫颈癌细胞的增殖、迁移和侵袭(P<0.05),抑制MiR-634的表达显著提高细胞的增殖、迁移和侵袭(P<0.05);抑制mTOR 的表达均显著降低细胞的增殖、迁移和侵袭(P<0.05)。结论:MiR-634 通过抑制mTOR 的表达来抑制宫颈癌细胞的增殖、迁移和侵袭,是宫颈癌基因靶向治疗的潜在靶点。     

Glucose-Regulated Protein 94 Mediates the Proliferation and Metastasis through the Regulation of ETV1 and MAPK Pathway in Colorectal Cancer

Colorectal cancer (CRC) is a worldwide health problem. Glucose-regulated protein 94 (GRP94) is known as an important endoplasmic reticulum-stress response protein that shows correlation with aggressive cancer behavior. However, the role of GRP94 in CRC is still unclear. Our results showed that silencing GRP94 (GRP94-KD) reduced cell proliferation, invasion and migration of CRC cells and suppressed tumorigenesis in the xenograft mouse model. Rescue assay showed that ETV1 overexpression reversed the effect of GRP94 on cell proliferation and migration. In the molecular mechanism, we found that knockdown of GRP94 inhibited the level of MAPK pathway, including ERK/p-ERK, JNK/p-JNK, and p38/p-p38 signals. Cyclooxygenase-2 and epithelial-mesenchymal transformation biomarkers, such as N-cadherin, vimentin, and β-catenin were suppressed in GRP94 knockdown cells. Treatment of specific inhibitors of MAPK pathway showed that ERK/p-ERK, and p38/p-p38 inhibitors significantly influenced ETV1 expression as compared to JNK/p-JNK inhibitor. Our results indicated that silencing GRP94 repressed the ability of EMT process, cancer cell proliferation, metastasis, and CRC tumorigenesis. Therefore, GRP94 may play an important role in CRC by regulating ETV1 and MAPK pathway.     

miR-21 inhibitor suppresses proliferation and migration of nasopharyngeal carcinoma cells through down-regulation of BCL2 expression

This study is to investigate the expression of miR-21 in nasopharyngeal carcinoma (NPC) cells, and the effect of miR-21 in the biological behavior and expression of B-cell lymphoma 2 (BCL2) in NPC cells. Paired NPC and adjacent non-tumor tissues were obtained from 53 patients who underwent primary surgical resection of NPC tissues. Luciferase reporter assay was performed to test whether BCL2 is a direct target of miR-21. Methylthiazolyl blue tetrazolium assay and colony assay were used to evaluate the effect of miR-21 on NPC cell proliferation. Transwell and wound-healing assays were carried out to test the effect of low expression of miR-21 on cancer cell migration and invasion. QRT-PCR and Western blotting were used to measure the levels of mRNA and protein expression, respectively. Tumor tissues showed a positive correlation between the levels of miR-21 and BCL2 protein expression. Cells transfected with miR-21 inhibitor healed slower compared the control (P < 0.05). In addition, cell migration was notably inhibited by the down-regulation of miR-21 in vitro (P < 0.05). The reduction in miR-21 expression showed a remarkable effect on the biological behavior of NPC cell clone formation (P < 0.05). Low expression of miR-21 by transfection with miRNA expression plasmid led to a decrease in BCL2 expression, which was accompanied by reduced migration and proliferation of the cancer cells. Our results demonstrated that miR-21 inhibitor down-regulated BCL2 expression level, suggesting that BCL2 might be a target gene for the initiation and development of NPC cells.