miR-124-3p靶向FOXQ1抑制宫颈鳞状细胞癌细胞增殖和侵袭的机制研究

目的:探讨miR-124-3p在宫颈鳞状细胞癌（cervical squamous cell carcinoma,CSCC）中的表达水平及对细胞增殖、侵袭能力的影响,初步阐明miR-124-3p对癌基因叉头框蛋白Q1(forkhead box Q1,FOXQ1)的靶向调控机制。方法:收集2015年1月至2017年12月手术切除的CSCC组织标本61份,分别应用RT-qPCR和IHC检测癌组织和癌旁组织中miR-124-3p和FOXQ1蛋白的表达水平,分析miR-124-3p和FOXQ1 mRNA表达的相关性;RT-qPCR检测miR-124-3p和FOXQ1 mRNA在人CSCC细胞株（SiHa和CaSki）及人宫颈永生化鳞状细胞株（Ect1/E6E7）中的表达水平;应用miR-124-3p模拟物转染CaSki细胞,分别采用CCK-8法和Transwell小室检测miR-124-3p对细胞增殖和侵袭能力的影响;采用Western blot检测miR-124-3p对FOXQ1、E-cadherin和Vimentin蛋白表达水平的影响;最后,应用双荧光素酶报告基因验证miR-124-3p对FOXQ1的靶向调控作用。结果:miR-124-3p在CSCC组织的表达水平低于癌旁组织（P＜0.05）,在人CSCC细胞株（SiHa和CaSki）中的表达水平也均低于人宫颈永生化鳞状细胞株（Ect1/E6E7）（P＜0.05）;与转染阴性对照细胞比较,转染miR-124-3p模拟物的CaSki细胞的增殖和侵袭能力明显受到抑制（P＜0.05）。FOXQ1 mRNA和蛋白在CSCC组织中的表达水平均显著高于癌旁组织(P＜0.05),相关性分析显示,FOXQ1 mRNA的表达水平与miR-124-3p呈负相关(r=-0.882,P＜0.05)。在转染miR-124-3p模拟物后,CSCC细胞株CaSki中FOXQ1和Vimentin蛋白表达水平均降低,而E-cadherin蛋白表达增高。双荧光素酶报告基因试验确认miR-124-3p可通过与FOXQ1 mRNA的3' UTR直接结合,靶向调控FOXQ1的表达。结论:miR-124-3p在CSCC中表达下调,其通过靶向调控FOXQ1的表达影响细胞的上皮间质转化状态,从而影响CSCC细胞的增殖和侵袭。     

Extracellular vesicles transmitted miR-31-5p promotes sorafenib resistance by targeting MLH1 in renal cell carcinoma

Sorafenib provides survival benefits in patients with advanced renal cell carcinoma (RCC), but its use is hampered by acquired drug resistance. It is important to fully clarify the molecular mechanisms of sorafenib resistance, which can help to avoid, delay or reverse drug resistance. Extracellular vesicles (EVs) can mediate intercellular communication by delivering effector molecules between cells. Here, we studied whether EVs are involved in sorafenib resistance of RCC and its possible molecular mechanisms. Using differential centrifugation, EVs were isolated from established sorafenib-resistant RCC cells (786-0 and ACHN), and EVs derived from sorafenib-resistant cells were uptaken by sensitive parental RCC cells and thus promoted drug resistance. Elevated exogenous miR-31-5p within EVs effectively downregulated MutL homolog 1 (MLH1) expression and thus promoted sorafenib resistance in vitro. Mice experiments also confirmed that miR-31-5p could mediate drug sensitivity in vivo. In addition, low expression of MLH1 was observed in sorafenib-resistant RCC cells and upregulation of MLH1 expression restored the sensitivity of resistant cell lines to sorafenib. Finally, miR-31-5p level in circulating EVs of RCC patients with progressive disease (PD) during sorafenib therapy was higher when compared to that in the pretherapy status. In conclusion, EVs shuttled miR-31-5p can transfer resistance information from sorafenib-resistant cells to sensitive cells by directly targeting MLH1, and thus magnify the drug resistance information to the whole tumor. Furthermore, miR-31-5p and MLH1 could be promising predictive biomarkers and therapeutic targets to prevent sorafenib resistance.     

miRNA profiling in metastatic renal cell carcinoma reveals a tumour-suppressor effect for miR-215

Background:

Renal cell carcinoma (RCC) is the most common neoplasm of the adult kidney. Metastatic RCC is difficult to treat. The 5-year survival rate for metastatic RCC is ⩽10%. Recently, microRNAs (miRNAs) have been shown to have a role in cancer metastasis and potential as prognostic biomarkers in cancer.

Method:

We performed a miRNA microarray to identify a miRNA signature characteristic of metastatic compared with primary RCCs. We validated our results by quantitative real-time PCR. We performed experimental and bioinformatic analyses to explore the involvement of miR-215 in RCC progression and metastasis.

Results:

We identified 65 miRNAs that were significantly altered in metastatic compared with primary RCCs. We validated our results by examining the expression of miR-10b, miR-126, miR-196a, miR-204 and miR-215, in two independent cohorts of patients. We showed that overexpression of miR-215 decreased cellular migration and invasion in an RCC cell line model. In addition, through gene expression profiling, we identified direct and indirect targets of miR-215 that can contribute to tumour metastasis.

Conclusion:

Our analysis showed that miRNAs are altered in metastatic RCCs and can contribute to kidney cancer metastasis through different biological processes. Dysregulated miRNAs represent potential prognostic biomarkers and may have therapeutic applications in kidney cancer.     

miR-124 and miR-506 inhibit colorectal cancer progression by targeting DNMT3B and DNMT1

miR-124 and miR-506 are reportedly down-regulated and associated with tumor progression in many cancers, but little is known about their intrinsic regulatory mechanisms in colorectal cancer (CRC). In this study, we found that the miR-124 and miR-506 levels were significantly lower in human CRC tissues than in controls, as indicated by qRT-PCR and in situ hybridization histochemistry. We also found that the overexpression of miR-124 or miR-506 inhibited tumor cell progression and increased sensitivity to chemotherapy in vitro. Increased miR-124 or miR-506 expression also inhibited tumor cell proliferation and invasion in vivo. Luciferase reporter assays and western blotting were used to determine the association between miR-124, miR-506 and their target genes, DNMTs. We further identified that miR-124 and miR-506 directly targeted DNMT3B and indirectly targeted DNMT1. The overexpression of miR-124 and miR-506 reduced global DNA methylation and restored the expression of E-cadherin, MGMT and P16. In conclusion, our data showed that miR-124 and miR-506 inhibit progression and increase sensitivity to chemotherapy by targeting DNMT3B and DNMT1 in CRC. These findings may provide novel avenues for the development of targeted therapies.     

miR-124通过调节Jagged1/Notch信号通路影响肾细胞癌OS-RC-2细胞的恶性生物学行为

目的：探讨miR-124通过调节Jagged1(JAG1)/Notch信号通路对肾细胞癌(RCC)细胞增殖、凋亡、迁移和侵袭的影响。方法：收集2018年6月至2021年10月在武汉市第三医院治疗的38例RCC患者的RCC组织和癌旁组织标本,并体外培养RCC细胞(Caki-2、A498、ACHN、786-O、OS-RC-2)和人正常肾细胞(293T),采用免疫组织化学法、qPCR和WB法检测miR-124和JAG1蛋白在RCC组织和细胞中的表达水平。选择miR-124表达与293T细胞差异最大的OS-RC-2细胞进行转染,按转染物不同分为Control组、NC mimic组、miR-124 mimic组、miR-124 mimic+pcDNA组和miR-124 mimic+pc-JAG1组。采用双荧光素酶报告基因实验验证miR-124与JAG1的关系;q PCR法检测miR-124、JAG1 mRNA表达;免疫组化法分析JAG1蛋白表达;WB法检测JAG1、凋亡相关蛋白(cleaved caspase-3、BAX和Bcl2)和Notch信号通路相关蛋白(NICD、HES1和HES5)的表...     

Downregulated miR-646 in clear cell renal carcinoma correlated with tumour metastasis by targeting the nin one binding protein (NOB1)

Background:

Nin one binding protein (NOB1) was identified as a potential oncogene in human glioma and miR-646 plays an important role in human growth and development. However, the underlying molecular mechanisms of NOB1 in tumorigenicity and its correlation with miR-646 in renal cell carcinoma (RCC) have not been investigated.

Methods:

We performed bioinformatic analysis to explore miRNA targeting NOB1. The expression of NOB1 and miR-646 from 100 cases of clear cell RCC (ccRCC) and 30 cases of adjacent non-tumour tissues were detected by quantitative real-time PCR. The expression of miR-646 was correlated with NOB1 expression, tumour features and patient metastasis-free survival. The effect of overexpression of mir-646 on renal cancer cell proliferation was detected by colony formation in soft agar. Using a xenograft tumour model, we observed the in vivo tumorigenesis effect of miR-646 and NOB1.

Results:

miR-646 negatively regulated NOB1 and inhibited the proliferation and migration of renal cancer cells. There was a significant upregulation of NOB1 in ccRCC and it was further increased in metastatic cases, while miR-646 was downregulated in tumour tissues and further decreased in metastatic ccRCC. Additionally, expression of miR-646 was inversely correlated with the expression of NOB1. The downregulation of miR-646 also indicated a higher probability of developing metastasis. Most importantly, miR-646 expression was an independent predictor of ccRCC metastasis by the univariate analysis and binary logistic regression model (both P<0.05). Colony formation in soft agar and xenograft tumour model suggested that miR-646 and NOB1 are required for tumorigenesis in vitro and in vivo. Furthermore, suppression of NOB1 increased the phosphorylation of several proteins in MAPK pathway.

Conclusions:

Downregulated miR-646 in ccRCC was associated with tumour metastasis through MAPK pathway by targeting NOB1. miR-646 and NOB1 may play an important role in the development of ccRCC.     

miR-124 通过靶向BECN1 基因调控细胞自噬抑制食管癌KYSE170 细胞的侵袭和迁移

目的：探讨miR-124 通过调控细胞自噬对食管癌KYSE170 细胞侵袭和迁移能力的影响。方法：食管癌KYSE170 细胞转染miR-124 mimic,Transwell 实验检测细胞侵袭和迁移能力的变化,双荧光素酶报告基因验证miR-124 对BECN1(Beclin1)基因的靶向调控作用,Western blotting 分析对BECN1、P62 及LC3 蛋白表达水平的影响。向KYSE170 细胞中转染BECN1 siRNA沉默BECN1 的表达,Transwell 法检测细胞侵袭及迁移能力的变化,Western blotting 检测BECN1、P62 及LC3 蛋白的表达。将miR-124 mimic 与BECN1 过表达质粒共转染至KYSE170 细胞,Transwell 实验检测细胞侵袭及迁移能力的变化,Western blotting 检测自噬相关基因表达的变化。结果：转染miR-124 mimic 后,KYSE170 细胞的侵袭和迁移能力下降(P<0.05),BECN1 蛋白及荧光素酶报告基因活性均明显下调(均P<0.01),自噬相关蛋白P62 表达增高,LC3 表达水平明显降低(均P<0.01)。沉默BECN1 表达抑制食管癌细胞侵袭及迁移(P<0.01),而过表达BECN1 使miR-124 mimic 对KYSE170 细胞自噬、侵袭和迁移能力的抑制作用明显减弱(P<0.01),自噬相关蛋白P62 表达降低、LC3 蛋白表达水平明显升高(均P<0.01)。结论：miR-124 能够抑制食管癌细胞侵袭及迁移能力,其机制可能与靶向调控自噬相关基因BECN1 的表达影响细胞自噬有关。     

lncRNA PTPRG-AS1通过靶向miR-124-3p调控肝癌细胞凋亡和放射敏感性的研究机制

目的:探讨lncRNA PTPRG-AS1对肝癌细胞凋亡和放射敏感性的影响及作用机制。方法:培养正常肝细胞L02和肝癌细胞HepG2、SMMC-7721和BEL-7402,qRT-PCR检测细胞中PTPRG-AS1和miR-124-3p表达水平。转染si-PTPRG-AS1、miR-124-3p mimics至HepG2细胞,抑制HepG2细胞中PTPRG-AS1表达或过表达miR-124-3p,流式细胞术检测细胞凋亡,克隆形成实验检测细胞放射敏感性,Western Blot法检测Bcl-2和Bax蛋白表达。生物信息学软件预测PTPRG-AS1与miR-124-3p存在互补的核苷酸序列,双荧光素酶报告基因实验验证PTPRG-AS1与miR-124-3p之间的调控关系。结果:与正常肝细胞L02相比,肝癌细胞HepG2、SMMC-7721和BEL-7402中PTPRG-AS1表达显著升高（P＜0.05）,miR-124-3p表达显著降低（P＜0.05）。抑制PTPRG-AS1或过表达miR-124-3p均可促进肝癌HepG2细胞的凋亡,增强肝癌HepG2细胞的放射敏感性,抑制Bcl-2蛋白表达,促进Bax蛋白表达。PTPRG-AS1负调控miR-124-3p表达。抑制miR-124-3p表达可部分逆转抑制PTPRG-AS1表达对肝癌HepG2细胞凋亡的促进作用以及放射敏感性的增强作用。结论:抑制PTPRG-AS1表达可能通过上调miR-124-3p表达促进肝癌细胞的凋亡并提高其放射敏感性,是肝癌治疗的潜在作用靶点。     

Downregulation of circ-TRPS1 suppressed prostatic cancer prognoses by regulating miR-124-3p/EZH2 axis-mediated stemness

Abnormal circular RNA (circRNA) expression correlates with human traits such as many kinds of cancers. Though circRNAs have links to cancer, they have less characterization in metastatic castration-resistant prostate cancer (PCa), which is main reason for PCa mortality. Therefore, high-throughput sequencing was used for selected circRNA profiles. The result showed that circ-TRPS1 was upregulated significantly in high-grade PCa tissues or cell lines. High circ-TRPS1 expression correlated to aggressive PCa phenotypes. Knockdown of circ-TRPS1 suppressed PCa proliferation and metastasis through targeting miR-124-3p/EZH2 axis-mediated stemness in PCa, which was validated by luciferase reporter assays. EZH2 overexpression or miR-124-3p inhibition reversed the inhibition of circ-TRPS1 silencing in PCa cell migration and proliferation by recovering stemness. In summary, data demonstrated that circ-TRPS1 suppressed PCa progression through functioning similar to a miR-124-3p sponge to enhance EZH2 expression and cancer stem-like cell differentiation. Thus, circ-TRPS1 might be a candidate target for PCa treatment.     

MiR-200c-3p inhibits cell migration and invasion of clear cell renal cell carcinoma via regulating SLC6A1

In this study, we investigated the mechanism of miR-200c-3p and SLC6A1 in regulating cell activity of clear cell renal cell carcinoma (CCRCC). The mRNA and miRNA expressions of tissue specimens were analyzed by CapitalBio Corporation (Beijing, China). The expression of SLC6A1 in CCRCC cells was examined through qRT-PCR and western blot. The migration and invasion ability of 786-O cells was testified by transwell assay after transfected. 786-O cell proliferation ability was detected by MTT assay. Dual luciferase reporter assay verified the association between SLC6A1 and miR-200c-3p. SLC6A1 was high expressed and miR-200c-3p was low expressed in CCRCC tissues and cells. Besides, lower SLC6A1 expression indicated longer survival time and higher survival rate. MiR-200c-3p could directly target at SLC6A1 and reduce its expression. MiR-200c-3p inhibited the proliferation, migration and invasion in 786-O cells by down-regulating SLC6A1 expression. The results suggested that the miR-200c-3p served as a suppressor for CCRCC via down-regulating SLC6A1.     

Chromosome 9p deletions identify an aggressive phenotype of clear cell renal cell carcinoma

BACKGROUND:

The authors investigated whether deletion of chromosome 9p in clear cell renal cell carcinoma (ccRCC) predicted worse disease‐specific survival (DSS) and recurrence‐free survival (RFS) and whether it was associated with more aggressive behavior in small renal masses.

METHODS:

In total, 703 ccRCC tumors were analyzed using fluorescence in situ hybridization (316 tumors) and cytogenetics (388 tumors). Tumor grade, classification, and size; 9p status; Eastern Cooperative Oncology Group performance status (ECOG PS); lymph node involvement; and the presence of metastasis were recorded. Outcomes were stratified by 9p status, and a Cox proportional hazards models was constructed using TNM staging, ECOG PS, tumor size, tumor grade, and 9p status.

RESULTS:

Deletions of 9p were detected in 97 tumors (13.8%). At presentation, 9p‐deleted tumors were larger and were more likely to be high grade (grade 3 or 4), to have a high tumor (T) classification (T3‐T4), and to have lymph node or distant metastases (P < .01). The median DSS for patients with and without 9p deletions was 37 months and 82 months, respectively (P < .01). In patients with localized disease, the median RFS in those who had 9p deletions was 53 months and was not reached in those without 9p deletions (P < .01). In patients who had localized lesions that measured ≤4 cm in greatest dimension, 9p‐deleted tumors were more likely to recur (19% vs 2%; P = .01).

CONCLUSIONS:

Deletion of chromosome 9p in ccRCC occurred in 14% of patients and was associated with higher grade and T classification, and the presence of lymph node and distant metastases. In addition, 9p deletion independently conferred a worse prognosis for patients with localized ccRCC, and most noteworthy, in patients with localized, small renal masses. Preoperatively identifying patients with 9p deletions will improve risk stratification and will help to select appropriate patients for surveillance protocols or aggressive treatment. Cancer 2010. © 2010 American Cancer Society.     

CircPVT1 promotes progression in clear cell renal cell carcinoma by sponging miR-145-5p and regulating TBX15 expression

Emerging evidence revealed that circular RNAs (circRNAs) play significant roles in regulating tumorigenesis and cancer progression. However, few circRNAs were well characterized in clear cell renal cell carcinoma (ccRCC). We found that circPVT1 was significantly upregulated in ccRCC tissues and positively associated with the clinical stage. The Area Under Curve of tissue and serum circPVT1 expression in ccRCC were 0.93 and 0.86, respectively. Importantly, we demonstrated that circPVT1 promoted ccRCC growth and metastasis in vitro and in vivo. We also found that circPVT1 directly binds to miRNA-145-5p via the Biotin-labelled miRNA pulldown assay and dual-luciferase reporter assay, and miR-145-5p inhibitor significantly attenuated the effect of circPVT1 knockdown on ccRCC cells. Moreover, through RNA sequencing and bioinformatics analysis, we demonstrated that TBX15 was regulated by the circPVT1/miR-145-5p axis and predicted poor prognosis in ccRCC. These findings suggest that circPVT1 promotes ccRCC growth and metastasis through sponging miR-145-5p and regulating downstream target TBX15 expression. The circPVT1/miR-145-5p/TBX15 axis might be a potential diagnostic marker and therapeutic target in ccRCC.     

miR-27a下调肾癌细胞FOXO1表达并促进其细胞增殖

目的 探讨microRNA-27a(miR-27a)在肾癌细胞中的作用及调控机制.方法 应用miR-27a反义寡核苷酸(ASO)在体外转染786-O和Caki-1细胞;qRT-PCR法检测miR-27a及FOXO1 mRNA的表达情况,Western blot检测FOXO1蛋白的表达水平;CCK8检测miR-27a ASO对细胞增殖的影响.结果 786-O和Caki-1细胞中miR-27a表达高于HK2正常肾小管上皮细胞(P<0.05);利用miR-27a ASO抑制786-O和Caki-1细胞miR-27a表达后,发现两个细胞株FOXO1 mRNA和蛋白表达的升高及增殖能力的降低(P<0.05).结论 miR-27a 可能通过调控FOXO1表达在肾癌中起致癌作用.miR-27a ASO可抑制肾癌细胞的增殖,因此miR-27a有可能作为肾癌基因治疗的候选靶点.     

miR-27a下调肾癌细胞FOXO1表达并促进其细胞增殖

目的：探讨microRNA-27a（miR-27a）在肾癌细胞中的作用及调控机制。方法：应用miR-27a反义寡核苷酸（ASO）在体外转染786-O和Caki-1细胞;qRT-PCR法检测miR-27a及FOXO1 mRNA的表达情况,Western blot检测FOXO1蛋白的表达水平;CCK8检测miR-27a ASO对细胞增殖的影响。结果：786-O和Caki-1细胞中miR-27a表达高于HK2正常肾小管上皮细胞（P〈0.05）;利用miR-27a ASO抑制786-O和Caki-1细胞miR-27a表达后,发现两个细胞株FOXO1 mRNA和蛋白表达的升高及增殖能力的降低（P〈0.05）。结论：miR-27a可能通过调控FOXO1表达在肾癌中起致癌作用。miR-27a ASO可抑制肾癌细胞的增殖,因此miR-27a有可能作为肾癌基因治疗的候选靶点。     

miR-1271-5p在肾细胞癌组织和细胞中的表达及其对A-498 细胞增殖及凋亡的影响

目的:探讨miR-1271-5p 在肾细胞癌（renal cell carcinoma,RCC）组织和细胞系中的表达及其对RCC细胞株A-498 增殖及凋亡的影响。方法：用实时荧光定量PCR（qPCR）检测手术切除并经病理确诊为RCC组织和癌旁组织,以及RCC细胞系ACHN、A498、HK-2、786-O、CaKi-1 和人胚肾细胞株HEK293 中miR-1271-5p 的表达水平。用miR-1271-5p（实验组）和miR-NC（对照组）分别转染A-498 细胞。通过生物信息学预测鸟嘌呤交换因子DOCK1 为miR-1271-5p 可能的靶基因,构建DOCK1 基因的3’UTR 野生型及突变体序列双荧光素酶报告基因载体并进行荧光素酶活性检测,qPCR检测两组细胞中DOCK1 mRNA表达水平,Western blotting 检测两组细胞中DOCK1、p-ERK、p-AKT、Bcl-2 和Bax蛋白的表达情况,MTS法、集落形成实验和流式细胞术检测A-489 细胞增殖、集落形成数目和凋亡情况。结果：RCC组织和细胞系中miR-1271-5p 表达水平显著低于癌旁组织和人胚肾HEK293 细胞（均P<0.01）。双荧光素酶报告基因系统结果显示DOCK1 是miR-1271-5p 的靶基因(P<0.01)。与miR-NC组细胞相比,miR-1271-5p 组A-498 细胞中DOCK1 mRNA的表达水平显著下降(P<0.01);DOCK1、p-ERK、p-AKT、Bcl-2 蛋白表达水平显著下调(P<0.05),Bax 蛋白明显上调(P<0.05);A-498 细胞增殖活力显著降低(P<0.01);集落形成数显著减少(P<0.05);细胞凋亡率显著增高(P<0.01)。结论：RCC组织和细胞系中miR-1271-5p 表达下调,通过干扰DOCK1 基因表达能明显抑制A-489 细胞的增殖及诱导其凋亡,miR-1271-5p 可能成为未来RCC治疗的分子靶标。     

lncRNA-NEAT1靶向miR-34 a抑制肾癌细胞生长的实验研究

目的:探讨长链非编码RNA-NEAT1(lncRNA-NEAT1)通过与miR-34a相互作用影响肾癌细胞生长的机制.方法:检测60例肾癌患者癌组织、癌旁组织中miR-34a及NEAT1表达水平的差异.稳定培养A498细胞,转染si-NEAT1,并用脂质体法瞬时转染miR-34a inhibitor,敲低miR-34a...     

miR-361-5p对肾细胞癌ACHN细胞恶性生物学行为的影响

目的：探讨miR-361-5p对肾细胞癌ACHN细胞增殖、侵袭、迁移、凋亡及其细胞周期的影响。方法：将miR-361-5p mimics和miR-361-5p inhibitor分别转染至肾癌ACHN细胞中,用qPCR检测转染细胞中miR-361-5p的表达水平,用MTT法、划痕愈合实验、Transwell实验、流式细胞术分别检测细胞的增殖、迁移、侵袭、细胞周期和凋亡水平。结果：与空白对照组和 Mimics-NC组比较,miR-361-5p mimics组ACHN细胞中miR-361-5p表达水平显著升高（P<0.01）,细胞的增殖、侵袭和迁移能力均显著减弱（均P<0.01）,而凋亡率升高（P<0.01）。与空白对照组或Inhibitor-NC组比较,miR-361-5p inhibitor组细胞中miR-361-5p表达水平显著下降（P<0.01）,细胞的增殖、侵袭和迁移能力均增强（均P<0.01）,细胞周期运转加速（P<0.01）,凋亡率降低（P<0.05）。结论：miR-361-5p可抑制肾癌ACHN细胞的增殖、侵袭和迁移,并诱导细胞凋亡,其在肾癌发生发展过程中发挥重要的抑制作用。     

CIAPIN1 inhibits the growth and proliferation of clear cell renal cell carcinoma

Our previous studies indicated a direct correlation with loss of CIAPIN1 and carcinogenesis of tumor in human gastric cancer. Here we presented that the expression of CIAPIN1 was absent or significantly decreased in 102 cases of clear cell renal cell carcinoma (CCRCC) tissues (P < 0.05). Up-regulating CIAPIN1 by adenoviral vectors exhibited significant inhibition of CCRCC-derived cell growth in vitro and in vivo with G1 cell cycle arrest. Simultaneously, CIAPIN1-induced growth suppression was found partially to regulate various proteins, including inhibition of cyclinD1, cyclinE, cdk2, cdk4, p-Rb and VEGF, but up-regulation of p27^Kip1 and Rb.     

miRNA-124-3p通过靶向STAT3抑制鼻咽癌细胞生长和转移

目的:探讨miR-124-3p在鼻咽癌中的表达及其对鼻咽癌细胞的影响及其机制.方法:收集鼻咽癌组织标本90例和慢性鼻咽炎症组织标本85例,运用qRT-PCR方法检测组织标本和CNE1、CNE2、SUNE1、HONE1、5-8F、6-10B及C666-1鼻咽癌细胞株和永生化鼻咽上皮细胞NP69中miR-124-3p的表达,脂质体介导的转染方法下调高表达miRNA-124-3p的鼻咽癌细胞株CNE2,同时上调低表达miRNA-124-3p的鼻咽癌细胞株C666-1.CCK8法、流式细胞技术、细胞划痕实验、Transwell实验和Boyden小室检测细胞增殖、凋亡、迁移及侵袭的变化;生物信息学靶基因预测miRNA-124-3的靶基因并运用荧光素酶报告实验验证,Western Blotting检测细胞中STAT3及其下游的p-STAT3、CCND2和MMP2蛋白表达水平.结果:鼻咽癌组织中miRNA-124-3p表达水平与慢性鼻咽炎症组织相比明显下调(P ＜0.001);miRNA-124-3p的表达水平与肿瘤大小及范围、区域淋巴结受累情况及临床分期显著相关(均P＜0.001);7种鼻咽癌细胞与NP69细胞相比较,miRNA-124-3p的表达水平均显著低于NP69(均P＜0.05).miRNA-124-3p过表达后C666-1细胞增殖、凋亡、迁移和侵袭均显著低于空白对照组和mimics NC组(均P＜0.05);miRNA-124-3p抑制后CNE2细胞的增殖、凋亡、迁移和侵袭均显著高于inhibitor NC组和空白对照组(均P＜0.05).转染miRNA-124-3p后C666-1细胞STAT3、p-STAT3、CCND2和MMP2的表达显著降低,抑制CNE2细胞的miRNA-124-3p表达后STAT3、p-STAT3、CCND2和MMP2的表达显著降低.结论:miR-124-3p可通过下调靶基因STAT3的表达,进而影响其下游的p-STAT3、CCND2和MMP2信号通路,从而促进鼻咽癌细胞凋亡,抑制鼻咽癌细胞的增殖、迁移及侵袭.