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Decreased long noncoding RNA SPRY4-IT1 contributing to gastric cancer cell metastasis partly via affecting epithelial-mesenchymal transition.

Xie M, Nie FQ, Sun M, Xia R, Liu YW, Zhou P, De W, Liu XH - J Transl Med (2015)

Bottom Line: Protein levels of SPRY4-IT1 targets were determined by western blot or fluorescence immunohistochemistry.Differences between groups were tested for significance using Student's t test (two-tailed).Patients with lower SPRY4-IT1 expression had a relatively poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 210029, Jiangsu, People's Republic of China. 1115769204@qq.com.

ABSTRACT

Background: Long noncoding RNAs (lncRNAs) are emerging as key regulators governing fundamental biological processes, and their disorder expression involves in tumorigenesis. SPRY4-IT1 (SPRY4 intronic transcript 1), a lncRNA derived from an intron within SPRY4 gene, involves in multiple cancers development. However, the expression pattern and biological function of SPRY4-IT1 in gastric cancer is still not well documented. Hence, we carried out the present study to investigate the potential role of SPRY4-IT1 in gastric carcinogenesis.

Methods: QRT-PCR was performed to detect the expression of SPRY4-IT1 in 61 pairs of gastric cancer samples. Over-expression and RNA interference (RNAi) approaches were used to investigate the biological functions of SPRY4-IT1. The effect of SPRY4-IT1 on proliferation was evaluated by MTT and colony formation assays. Gastric cancer cells transfected with pCDNA-SPRY4-IT1 were injected into nude mice to study the effect of SPRY4-IT1 on tumorigenesis and metastasis in vivo. Protein levels of SPRY4-IT1 targets were determined by western blot or fluorescence immunohistochemistry. ChIP assays were performed to investigate the effect of DNMT1 on SPRY4-IT1 expression. Differences between groups were tested for significance using Student's t test (two-tailed).

Results: SPRY4-IT1 expression is decreased in gastric cancer tissues and associated with larger tumor size, advanced pathological stage, deeper depth of invasion and lymphatic metastasis. Patients with lower SPRY4-IT1 expression had a relatively poor prognosis. DNA methylation may be a key factor in controlling the SPRY4-IT1 expression. Furthermore, SPRY4-IT1 contributed to gastric cancer cells metastasis might partly via regulating epithelial-mesenchymal transition (EMT) process.

Conclusion: Low expression of SPRY4-IT1 is involved in progression and metastasis of gastric cancer and may represent a novel biomarker of poor prognosis in patients with gastric cancer.

No MeSH data available.


Related in: MedlinePlus

The effect of SPRY4-IT1 on gastric cancer cells proliferation in vitro and vivo. a MTT assay was performed to determine the proliferation of pCDNA-SPRY4-IT1 or si-SPRY4-IT1 transfected BGC823 and SGC7901 cells. Data represent the mean ± SD from three independent experiments. b Colony-forming growth assay was performed to determine the colony formation ability of pCDNA-SPRY4-IT1 transfected BGC823 and SGC7901 cells. The colonies were counted and captured. c The tumor volume was calculated every 3 days after injection of BGC823 cells stably transfected with pCDNA-SPRY4-IT1 or empty vector. d Tumor weights are represented as means of tumor weights ± SD. e Tumors developed from pCDNA-SPRY4-IT1 transfected BGC823 cells showed lower ki67 protein levels than tumors developed by control cells. Upper H & E staining; lower: immunostaining. *P < 0.05 and **P < 0.01.
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Fig3: The effect of SPRY4-IT1 on gastric cancer cells proliferation in vitro and vivo. a MTT assay was performed to determine the proliferation of pCDNA-SPRY4-IT1 or si-SPRY4-IT1 transfected BGC823 and SGC7901 cells. Data represent the mean ± SD from three independent experiments. b Colony-forming growth assay was performed to determine the colony formation ability of pCDNA-SPRY4-IT1 transfected BGC823 and SGC7901 cells. The colonies were counted and captured. c The tumor volume was calculated every 3 days after injection of BGC823 cells stably transfected with pCDNA-SPRY4-IT1 or empty vector. d Tumor weights are represented as means of tumor weights ± SD. e Tumors developed from pCDNA-SPRY4-IT1 transfected BGC823 cells showed lower ki67 protein levels than tumors developed by control cells. Upper H & E staining; lower: immunostaining. *P < 0.05 and **P < 0.01.

Mentions: To assess the biological role of SPRY4-IT1 in gastric cancer, we firstly investigated the effect of over-expression of SPRY4-IT1 on cell proliferation. MTT assays showed that cell growth was significantly impaired in pCDNA-SPRY4-IT1 transfected SGC7901 cells or BGC823 cells (Fig. 3a), while knockdown of SPRY4-IT1 expression promoted BGC823 cells proliferation (Additional file 2: Figure S1B). Similarly, the results of colony-formation assays showed that clonogenic survival was decreased following SPRY4-IT1 over-expression in SGC7901 cells or BGC823 cells (Fig. 3b).Fig. 3


Decreased long noncoding RNA SPRY4-IT1 contributing to gastric cancer cell metastasis partly via affecting epithelial-mesenchymal transition.

Xie M, Nie FQ, Sun M, Xia R, Liu YW, Zhou P, De W, Liu XH - J Transl Med (2015)

The effect of SPRY4-IT1 on gastric cancer cells proliferation in vitro and vivo. a MTT assay was performed to determine the proliferation of pCDNA-SPRY4-IT1 or si-SPRY4-IT1 transfected BGC823 and SGC7901 cells. Data represent the mean ± SD from three independent experiments. b Colony-forming growth assay was performed to determine the colony formation ability of pCDNA-SPRY4-IT1 transfected BGC823 and SGC7901 cells. The colonies were counted and captured. c The tumor volume was calculated every 3 days after injection of BGC823 cells stably transfected with pCDNA-SPRY4-IT1 or empty vector. d Tumor weights are represented as means of tumor weights ± SD. e Tumors developed from pCDNA-SPRY4-IT1 transfected BGC823 cells showed lower ki67 protein levels than tumors developed by control cells. Upper H & E staining; lower: immunostaining. *P < 0.05 and **P < 0.01.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4522960&req=5

Fig3: The effect of SPRY4-IT1 on gastric cancer cells proliferation in vitro and vivo. a MTT assay was performed to determine the proliferation of pCDNA-SPRY4-IT1 or si-SPRY4-IT1 transfected BGC823 and SGC7901 cells. Data represent the mean ± SD from three independent experiments. b Colony-forming growth assay was performed to determine the colony formation ability of pCDNA-SPRY4-IT1 transfected BGC823 and SGC7901 cells. The colonies were counted and captured. c The tumor volume was calculated every 3 days after injection of BGC823 cells stably transfected with pCDNA-SPRY4-IT1 or empty vector. d Tumor weights are represented as means of tumor weights ± SD. e Tumors developed from pCDNA-SPRY4-IT1 transfected BGC823 cells showed lower ki67 protein levels than tumors developed by control cells. Upper H & E staining; lower: immunostaining. *P < 0.05 and **P < 0.01.
Mentions: To assess the biological role of SPRY4-IT1 in gastric cancer, we firstly investigated the effect of over-expression of SPRY4-IT1 on cell proliferation. MTT assays showed that cell growth was significantly impaired in pCDNA-SPRY4-IT1 transfected SGC7901 cells or BGC823 cells (Fig. 3a), while knockdown of SPRY4-IT1 expression promoted BGC823 cells proliferation (Additional file 2: Figure S1B). Similarly, the results of colony-formation assays showed that clonogenic survival was decreased following SPRY4-IT1 over-expression in SGC7901 cells or BGC823 cells (Fig. 3b).Fig. 3

Bottom Line: Protein levels of SPRY4-IT1 targets were determined by western blot or fluorescence immunohistochemistry.Differences between groups were tested for significance using Student's t test (two-tailed).Patients with lower SPRY4-IT1 expression had a relatively poor prognosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Nanjing Medical University, Nanjing, 210029, Jiangsu, People's Republic of China. 1115769204@qq.com.

ABSTRACT

Background: Long noncoding RNAs (lncRNAs) are emerging as key regulators governing fundamental biological processes, and their disorder expression involves in tumorigenesis. SPRY4-IT1 (SPRY4 intronic transcript 1), a lncRNA derived from an intron within SPRY4 gene, involves in multiple cancers development. However, the expression pattern and biological function of SPRY4-IT1 in gastric cancer is still not well documented. Hence, we carried out the present study to investigate the potential role of SPRY4-IT1 in gastric carcinogenesis.

Methods: QRT-PCR was performed to detect the expression of SPRY4-IT1 in 61 pairs of gastric cancer samples. Over-expression and RNA interference (RNAi) approaches were used to investigate the biological functions of SPRY4-IT1. The effect of SPRY4-IT1 on proliferation was evaluated by MTT and colony formation assays. Gastric cancer cells transfected with pCDNA-SPRY4-IT1 were injected into nude mice to study the effect of SPRY4-IT1 on tumorigenesis and metastasis in vivo. Protein levels of SPRY4-IT1 targets were determined by western blot or fluorescence immunohistochemistry. ChIP assays were performed to investigate the effect of DNMT1 on SPRY4-IT1 expression. Differences between groups were tested for significance using Student's t test (two-tailed).

Results: SPRY4-IT1 expression is decreased in gastric cancer tissues and associated with larger tumor size, advanced pathological stage, deeper depth of invasion and lymphatic metastasis. Patients with lower SPRY4-IT1 expression had a relatively poor prognosis. DNA methylation may be a key factor in controlling the SPRY4-IT1 expression. Furthermore, SPRY4-IT1 contributed to gastric cancer cells metastasis might partly via regulating epithelial-mesenchymal transition (EMT) process.

Conclusion: Low expression of SPRY4-IT1 is involved in progression and metastasis of gastric cancer and may represent a novel biomarker of poor prognosis in patients with gastric cancer.

No MeSH data available.


Related in: MedlinePlus