Weiliang Sun

Long non-coding RNA expression profile in human gastric cancer and its clinical significances.

BackgroundLong non-coding RNAs (lncRNAs) are prevalently transcribed in the genome yet their potential roles in human cancers are not well understood. The aim of the present study was to determine the lncRNA expression profile in gastric cancer and its potential clinical value.MethodsThe global lncRNA expression profile in gastric cancer was measured by lncRNA microarray. Levels of two representative lncRNAs, H19 and uc001lsz, were confirmed by real-time reverse transcriptase-polymerase chain reaction. The relationship between their levels and clinicopathological factors of patients with gastric cancer was explored. A receiver operating characteristic (ROC) curve was constructed for differentiating gastric cancer from benign gastric diseases.ResultsTotal of 135 lncRNAs, which differential expression levels between tumor and non-tumorous tissues were more than twofold, were found (GEO No. GSE47850). The most down-regulated lncRNAs in gastric cancer tissues were FER1L4, uc001lsz, BG491697, AF131784, uc009ycs, BG981369, AF147447, HMlincRNA1600, and AK054588; while the most up-regulated ones were H19, HMlincRNA717, BM709340, BQ213083, AK054978, and DB077273. H19 was found highly expressed in stomach and liver cancer cell lines, while lowly expressed in lung cancer and prostate cancer cell lines. Uc001lsz was lowly expressed in gastric, lung and liver cancer cell lines, while highly expressed in prostate cancer. The areas under ROC curves were up to 0.613, 0.751, and 0.761 for H19, uc001lsz, and the combination, respectively.ConclusionsThe lncRNA expression profile in gastric cancer suggests the potential roles of lncRNAs in gastric cancer occurrence and development. The overexpression of H19 in gastric cancer suggests that H19 may be participated in gastric cancer. The reduced expression of uc001lsz in gastric cancer cell lines and tissues, its associations with TNM stage, and its dysregulation in early cancer and precancerous lesions suggest that uc001lsz may be a potential marker for the diagnosis of early gastric cancer.

MicroRNA-195 and microRNA-378 mediate tumor growth suppression by epigenetical regulation in gastric cancer

The epigenetic regulation of microRNAs is one of several mechanisms underlying carcinogenesis. We found that microRNA-195 (miR-195) and microRNA-378 (miR-378) were significantly down-regulated in gastric cancer tissues and gastric cancer cell lines. The expression of miR-195 and miR-378 in gastric cancer cells was significantly restored by 5-aza-dC, a demethylation reagent. The low expression of miR-195 and miR-378 was closely related to the presence of promoter CpG island methylation. Treatment with miR-195/miR-378 mimics strikingly suppressed the growth of gastric cancer cells whereas promoted the growth of normal gastric epithelial cells. In contrast, administration of miR-195/miR-378 inhibitors significantly prevented the growth of normal gastric epithelial cells. Expression of cyclin-dependent kinase 6 and vascular endothelial growth factor was down-regulated by exogenous miR-195 and miR-378, respectively. In conclusion, miR-195 and miR-378 are abnormally expressed and epigenetically regulated in gastric cancer cell lines and tissues via the suppression of CDK6 and VEGF signaling, suggesting that miR-195 and miR-378 have tumor suppressor properties in gastric cancer.

Gastric juice long noncoding RNA used as a tumor marker for screening gastric cancer

Long noncoding RNAs (lncRNAs) play a crucial role in tumorigenesis. However, the value of lncRNAs in the diagnosis of gastric cancer remains unknown. To identify whether lncRNA‐AA174084 is a potential marker for the early diagnosis of gastric cancer (GC), the authors investigated its levels in tissues, blood, and gastric juices from patients with various stage of gastric tumorigenesis.

Decreased expression of long noncoding RNA AC096655.1-002 in gastric cancer and its clinical significance.

Long noncoding RNAs (lncRNAs) are newfound noncoding RNAs that are greater than 200 nucleotides in length. They have emerged recently as major players in governing fundamental biological processes. However, the expression level of lncRNAs and their clinical significances are not well understood. To investigate the lncRNA expression in gastric cancer, real-time reverse transcriptase-polymerase chain reaction was conducted. Then, the association between the level of AC096655.1-002, one of lncRNA, in gastric cancer tissues and the clinicopathological features of patients with gastric cancer was further analyzed. Receiver operating characteristic (ROC) curve was constructed to evaluate the diagnostic values. The results showed that AC096655.1-002 was significantly downregulated in gastric cancer tissues compared with paired adjacent non-tumorous tissues (P < 0.001). Its expression level was significantly correlated with lymph node metastasis (P < 0.001), distant metastasis (P < 0.001), tumor–node–metastasis stages (P < 0.001), and differentiation (P = 0.030). The area under the ROC curve of AC096655.1-002 was up to 0.731. For the detection of gastric cancer, the use of AC096655.1-002 showed a remarkable improvement compared with the use of serum carcinoembryonic antigen. These results indicated that lncRNA AC096655.1-002 may be a potential biomarker in the diagnosis of gastric carcinoma.

MicroRNA-21 is a new marker of circulating tumor cells in gastric cancer patients

The detection of circulating tumor cells (CTCs) has received great attention. MicroRNA-21 (miR-21) plays crucial roles in carcinogenesis and is considered as one of the most studied oncomiRNAs. We determined if miR-21 could be used a marker for the detection of CTCs in gastric cancer patients. Peripheral blood samples were collected from 53 preoperative patients with gastric cancer and 20 healthy volunteers. Real-time reverse transcription-polymerase chain reaction was used to detect the level of miR-21. Receiver operator characteristic curves (ROC) were constructed. Patients with gastric cancer display a significantly higher level of miR-21 in peripheral blood than those from controls. The miR-21 level was associated with the tumor node metastasis (TNM) stage, tumor size and tissue categories. The area under ROC curve was up to 0.853 ± 0.086. This study highlights the potential of the detection of miR-21 in peripheral blood as a novel tool for monitoring CTCs in gastric cancer patients.

LncRNA-RMRP promotes carcinogenesis by acting as a miR-206 sponge and is used as a novel biomarker for gastric cancer

Long noncoding RNAs (lncRNAs) play crucial roles in tumorigenesis. However, the mechanisms of most lncRNAs in cancers are largely unknown. Because the RNA component of mitochondrial RNA processing endoribonuclease (RMRP) is one of the dysregulated lncRNAs in gastric cancer, this study explored its molecular mechanisms in carcinogenesis. RMRP levels in 792 tissues, plasma and gastric juices from patients with various stages of gastric tumorigenesis were analyzed by quantitative reverse transcription-polymerase chain reaction. Overexpression and RNA interference were used to manipulate RMRP expression by RMRP expression vector and small interfering RNAs, respectively. Its mechanisms were evaluated by flow cytometry, real-time cell analysis, plate colony formation assays, and xenograft models. RMRP levels in tissue, plasma and gastric juices from patients with gastric cancer were significantly different from those from controls. Its levels were significantly associated with Borrmann type and metastasis. Plasma and gastric juice RMRP had higher sensitivity and specificity than commonly used markers (such as carcinoembryonic antigen and carbohydrate antigen 19–9). Knockdown of RMRP significantly inhibited cell proliferation in vitro and in vivo, whereas overexpression of RMRP promoted cell growth. Acting as a miR-206 sponge, RMRP modulated cell cycle by regulating Cyclin D2 expression. RMRP plays a crucial role in gastric cancer occurrence and can be used as a novel biomarker for gastric cancer.

lncRNA-AC130710 targeting by miR-129-5p is upregulated in gastric cancer and associates with poor prognosis

Long noncoding RNAs (lncRNAs) play an important role in cancer occurrence and development. However, there is largely unknown about lncRNAs significance in the diagnosis and treatment of gastric cancer. In our study, we focused on AC130710, one of lncRNAs. Gastric cancer tissues and adjacent tissues were gathered from 78 patients with gastric cancer. The AC130710 levels were detected by reverse transcription polymerase chain reaction (RT-PCR). Then, we further analyzed the association between AC130710 level and the clinicopathological factors of patients with gastric cancer. Finally, the molecular mechanism underling AC130710 highly expressed in gastric cancer cells was explored. The results showed that AC130710 in cancer tissues from patients with gastric cancer was significantly higher than those in adjacent noncancerous tissues (P < 0.05). Its expression level was significantly associated with tumor size (P = 0.013), tumor-node-metastasis (TNM) stages (P = 0.030), and distal metastasis (P = 0.018). AC130710 expression in MGC-803 was significantly higher than that in normal gastric mucosa cell line GES-1 (P < 0.001). Moreover, miR-129-5p may play an important role in the downregulation of AC130710 in gastric cancer cells. These results indicated that lncRNA-AC130710 may be a potential tumor marker for gastric cancer prognosis.

Regulatory mechanisms of long noncoding RNAs on gene expression in cancers.

Long non-coding RNAs (lncRNAs) are a heterogeneous class of RNAs that are non-protein coding transcripts longer than 200 nucleotides. In this review, we introduce the mechanisms by which lncRNAs regulate gene expression in four parts, epigenetic regulation (genetic imprinting and chromatin remodeling), transcriptional regulation (molecular decoy), post-transcriptional regulation (splicing and mRNA decay), and translational regulation. H19, Xist, and others are involved in genomic imprinting. HOTAIR and ANRIL function in chromatin remodeling. GAS5 is degraded through an RNA decay pathway. NEAT1 and MALAT1 function not only in the regulation of transcription but also in splicing.

Roles of long noncoding RNAs in gastric cancer and their clinical applications.

PurposeGastric cancer ranks as the most common cancer in the world. However, the progresses of its diagnosis and treatment are still not satisfactory. The purpose of this study is to summarize the roles of lncRNAs associated with gastric cancer.MethodsWe searched lncRNAs associated with gastric cancer in PubMed.ResultsLong noncoding RNAs (lncRNAs), transcripts larger than 200 nucleotides, regulate gene expression at various levels. They are playing important roles in the occurrence and development of gastric cancer. They are involved in signaling pathways, crosstalk with microRNAs, and affecting metastasis by regulating epithelial-to-mesenchymal transition. By acting as oncogenes or tumor suppressors, lncRNAs contribute to gastric cancer occurrence and development. Several lncRNAs including HOTAIR, HULC, LINC00152, MALAT2, H19, GHET1, and GACAT3 have been demonstrated having oncogene activities, while other lncRNAs including LEIGC, GAS5, and FER1L4 have been thought as tumor suppressors.ConclusionsSeveral lncRNAs from tissue, blood, and gastric juice have shown potential values in gastric cancer diagnosis or prognosis evaluation.

Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection

To observe the diagnostic values of circular RNAs (circRNAs), their expression profiles between gastric cancer patients’ plasma and healthy controls were first assessed by circRNA microarray. Then, circRNA levels were measured by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and RT-droplet digital PCR (RT-ddPCR), respectively. A total of 343 differentially expressed circRNAs were found. The top 10 elevated circRNAs in patients were hsa_circ_0088300, hsa_circ_0075825, hsa_circ_0019172, hsa_circ_0000220, hsa_circ_0035277, hsa_circ_0000301, hsa_circ_0000189, hsa_circ_0090080, hsa_circ_0001888, and hsa_circ_0001874. The top 10 reduced circRNAs were hsa_circ_0004771, hsa_circ_0001190, hsa_circ_0061276, hsa_circ_0092337, hsa_circ_0058495, hsa_circ_0061274, hsa_circ_0075829, hsa_circ_0080845, hsa_circ_0001006, and hsa_circ_0003764. In cancer and dysplasia tissues, hsa_circ_0001017 and hsa_circ_0061276 were downregulated. Their levels were significantly associated with distal metastasis. The area under receiver operating characteristic curve in combinative use was 0.966 with 95.5% sensitivity and 95.7% specificity. Patients with low plasma hsa_circ_0001017 or hsa_circ_0061276 had a much shorter overall survival than those with high levels. Patients whose plasma hsa_circ_0001017 or hsa_circ_0061276 levels recovered to normal after operation had a longer disease-free survival. Finally, the in vitro model indicated gastric cancer cells secreting circRNAs into plasma. In conclusion, RT-ddPCR is a potent non-invasive and absolute quantification method for simultaneous detection of multiple circRNAs. Hsa_circ_0001017 and hsa_circ_0061276 are new potential biomarkers for gastric cancer.Key messageA total of 343 circRNAs are differentially expressed between gastric cancer patients’ plasma and healthy controls.Hsa_circ_0001017 and hsa_circ_0061276 are downregulated in gastric cancer tissues.The RT-ddPCR is a potent method for simultaneous detection of multiple circRNAs in plasma.Hsa_circ_0001017 and hsa_circ_0061276 are potential biomarkers for gastric cancer.