Study of long non-coding RNA Tug 1 expression in Egyptian colorectal adenocarcinoma patients

Purpose: Colorectal cancer (CRC) is one of the most fatal tumors worldwide. In Egypt, most CRC cases occur in individuals > 40 years old. TUG1 has been proved to be disrupted in different malignancies and may have a critical role in tumor progression, invasion, and metastasis. However, its role in CRC has not been adequately studied. Materials / Methods: Quantitative real-time polymerase chain reaction (PCR) was used to evaluate the expression levels of long non-coding RNA (LncRNA) taurine upregulated gene 1 (TUG1), in nonmetastatic and metastatic CRC tissues and adjacent noncancerous tissues as control. Results: LncRNA TUG1 expression was significantly upregulated in both nonmetastatic and metastatic CRC tissues, in comparison with the adjacent noncancerous tissue. It was found that TUG1 could have a possible prognostic role in CRC, by comparing the sensitivity and specificity of TUG1 with those of CEA and CA19-9. Conclusion: The results of the current study suggest that the LncRNA TUG1 participates in the malignant behaviors of CRC cells. (AU)

Mogroside v stimulates osteoblast proliferation and differentiation by promoting lncRNA TUG1 expression / 中国组织工程研究

BACKGROUND: Osteoporosis is a balance disorder between bone formation of osteoblasts and bone resorption of osteoclasts during bone remodeling. Strict control of bone remodeling at the cellular level is important to maintain bone homeostasis and avoid osteoporosis. Previous studies have shown that 1.25×10-2 g/L mogroside V can promote osteoblast proliferation and differentiation, and its mechanism may be related to LncRNA TUG1. OBJECTIVE: To investigate the role of LncRNA TUG1 in the promotion of osteoblast proliferation and differentiation by mogroside V. METHODS: Osteoblasts from neonatal Sprague-Dawley rats were extracted by two-step enzymatic digestion. The cells were divided into two groups and treated with 0 and 1.25×10-2 g/L Mogroside V. The LncRNA was detected after 2 days of culture. LncRNA TUG1 silencing virus was designed and synthetized. The newly extracted osteoblasts were divided into normal cell control group, mogroside V intervention group, mogroside V+negative virus group, TUG1 silent group, and mogroside V+TUG1 silent group. The proliferation of osteoblasts was observed by FDA fluorescence staining at 2, 4, and 6 days after processing according to the above grouping conditions. After 6 days of treatment on osteoblasts, the effect of TUG1 on osteoblast proliferation and differentiation was studied by alkaline phosphatase staining, alizarin red staining and qRT-PCR. RESULTS AND CONCLUSION: LncRNA detection showed that 1.25×10-2 g/L Mogroside V significantly promoted the expression of LncRNA TUG1 in osteoblasts. FDA fluorescent staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on osteoblast proliferation. After 6 days of treatment, alkaline phosphatase staining and alizarin red staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on mineralization of osteoblasts. The results of qRT-PCR showed that Runx2, BSP, OCN and COL1A1 genes were highly expressed in the mogroside V intervention group, but their expression was inhibited in the mogroside V+TUG1 silent group. Overall findings indicate that mogroside V stimulates the proliferation and differentiation of osteoblasts by promoting the expression of LncRNA TUG1.

Knockdown of lncRNA TUG1 inhibits hippocampal neuronal apoptosis and participates in aerobic exercise-alleviated vascular cognitive impairment

OBJECTIVES: Our previous study indicated that aerobic exercise relieves cognitive impairment in patients with vascular cognitive impairment (VCI) via regulating brain-derived neurotrophic factor (BDNF), but the mechanism is not yet clear. This study aimed to explore whether lncRNA taurine upregulated gene 1 (TUG1) participates in the process of VCI by regulating BDNF. METHODS: The expressions of TUG1 and BDNF in the serum of VCI patients were detected. The potential molecular mechanisms of TUG1 in regulating hippocampal neuronal apoptosis were explored in oxygen and glucose deprivation-induced (OGD-induced) hippocampal cell line HT22. The VCI mouse model was established, and TUG1 and BDNF were overexpressed via lentivirus injection. The cognitive impairment of mice was detected by the Morris water maze experiment after the aerobic exercise. RESULTS: The level of TUG1 was elevated in the serum of VCI patients compared with the control group. The knockdown of TUG1 in OGD-induced HT22 cells increased BDNF level and decreased cell apoptosis, and the downregulation of BDNF restored the decreased cell apoptosis. RNA immunoprecipitation and RNA pull-down assays showed that TUG1 could bind to BDNF protein. The aerobic exercise alleviated cognitive impairment and inhibited hippocampal apoptosis in VCI mice. Meanwhile, the overexpression of TUG1 reversed the therapeutic effects of aerobic exercise on cognitive impairment. CONCLUSIONS: The knockdown of TUG1 reduced hippocampal neuronal apoptosis and participates in the aerobic exercise-alleviated VCI, which was partly through regulating BDNF.

lncRNA TUG1 promotes proliferation and differentiation of osteoblasts by regulating the miR-545-3p/CNR2 axis

Osteoblast differentiation is an effective way to promote bone formation. Long non-coding RNA taurine upregulated 1 (TUG1) has been identified as a crucial modulator of multiple biological processes. This study was designed to investigate the function of TUG1 in the proliferation and differentiation of osteoblast precursor cells hFOB1.19. In this study, we found that TUG1 promoted hFOB1.19 cell proliferation, while TUG1 knockdown hindered cell proliferation. TUG1 and cannabinoid receptor 2 (CNR2) were upregulated, while miR-545-3p was down-regulated in hFOB1.19 cells undergoing osteoblastic differentiation. TUG1 induced osteoblast differentiation by increasing alkaline phosphatase (ALP) activity and the expression of osteoblastic differentiation markers. TUG1 was a sponge of miR-545-3p and regulated osteoblastic differentiation by modulating miR-545-3p. Moreover, miR-545-3p directly targeted CNR2 and restored the effect of CNR2 on osteoblastic differentiation. In conclusion, TUG1 accelerated the proliferation and differentiation of osteoblasts by sponging miR-545-3p and increasing CNR2 expression, which might provide a new biomarker for bone diseases.

Mir-29c-3p targeting TUG1 affects migration and invasion of bladder cancer cells by regulating CAPN7 expression / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which long non-coding RNA TUG1 affects bladder cancer cell migration and invasion.@*METHODS@#The expressions of TUG1 and miR-29c-3p were examined by quantitative RT-PCR (qRT-PCR) in 10 bladder cancer tissues and 5 bladder cancer cell lines. Trans-well assay was used to detect the changes in migration and invasion abilities of bladder cancer T24 cells after TUG1 knockdown using RNA interference technique, and the alteration in the expression of CAPN7 was also detected. The expression of CAPN7 was examined in T24 cells overexpressing mir-29c-3p by Western blotting, and luciferase reporter assay was performed to confirm the targeting of miR-29c-3p to TUG1 and CAPN7. The effects of CAPN7 overexpression and sh-TUG1 on the migration and invasion of T24 cells were investigated.@*RESULTS@#The expression of TUG1 was up-regulated and mir-29c-3p was down-regulated significantly in bladder cancer tissue with a negative correlation between their expressions. TUG1 knockdown significantly inhibited the migration and invasion of T24 cells ( < 0.01). Overexpression of mir-29c-3p in T24 cells obviously down-regulated the expression of CAPN7 protein, whose expression was positively correlated with TUG1 expression (=0.4081, =0.0139). The results of luciferase reporter assay confirmed both TUG1 and CAPN7 as the targets of mir-29c-3p. CAPN7 overexpression could partially reverse the tumor suppressing effect of sh-TUG1 in T24 cells.@*CONCLUSIONS@#Mir-29c-3p targeting TUG1 affects the migration and invasion of bladder cancer cells by regulating the expression of CAPN7.

Effects of long non-coding RNA TUG1 on the proliferation and apoptosis of gastric cancerAGS cells / 中国肿瘤生物治疗杂志

@#Objective: To investigate the expression of lncRNA TUG1 (long non-coding RNA taurine up-regulated gene 1) in gastric cancer and its effect on the proliferation and apoptosis of gastric cancer cells. Methods: Surgically resected gastric cancer tissues and corresponding distal normal tissues (>5 cm away from the margin of tumor) of 40 gastric cancer patients from March 2016 to December 2017 at Ganzhou People's Hospital of Jiangxi Province were collected, and qPCR was used to examine the expression of lncRNA TUG1.AGS gastric cancer cells were transfected with lncRNATUG1 over-expression plasmids and siRNAs, and the effects of lncRNA TUG1 on cell proliferation and apoptosis were assessed by CCK-8, qPCR and Flow cytometry. Results: lncRNATUG1 expression was significantly increased in gastric cancer tissues as compared to normal tissues; and it was not correlated with gender, age, tumor size, infiltration depth of tumor, lymph node-metastasis, tumor differentiation and TNM staging. TUG1 over-expression significantly suppressed the expressions of CDKN1A, BAX and Caspase-3 in AGS gastric cancer cells, and decreased G1 phase proportion and apoptosis rate, but increased S phase proportion and cell viability; in contrast, TUG1 siRNA transfection significantly promoted the expressions of CDKN1A, BAX and Caspase-3, and increased G1 phase proportion and apoptosis rate, but decreased S phase proportion and cell viability. Conclusion: Up-regulated lncRNATUG1 promotes proliferation and inhibits apoptosis of gastric cancer cells.

Effect of long-chain non-coding RNA TUG1 on radiosensitivity of human cervical cancer XB1702 cells by adsorption of miR-145 / 中华放射肿瘤学杂志

Objective@#To evaluate the effect of long-chain non-coding RNA TUG1 on the radiosensitivity of cervical cancer cells and explore its underlying mechanism.@*Methods@#The expression of TUG1 and miR-145 in cervical cancer cells XB1702 and normal endometrial stromal cells (ESCs) was detected by qRT-PCR. The transfected si-NC, transfected si-TUG1, transfected si-NC combined with irradiation, transfected si-TUG1 combined with irradiation, si-TUG1 and anti-miR-NC co-transfected and, si-TUG1 and anti-miR-145 co-transfected groups were established, which were transfected into XB1702 cells by liposome method. The survival fraction of each group was detected by colony formation assay. The cell apoptosis of each group was detected by flow cytometry. The fluorescence activity of each group was assessed by dual luciferase reporter gene assay.@*Results@#Compared with the normal ESCs, the expression of TUG1 was significantly up-regulated, whereas that of miR-145 was significantly down-regulated in the cervical cancer cells XB1702. Silencing TUG1 significantly increased the survival fraction of XB1702 cells, promoted cell apoptosis and enhanced the radiosensitivity of irradiation to XB1702 cells. TUG1 could target and regulate the expression of miR-145. Suppressing miR-145 reversed the silencing effect of TUG1 on inhibiting proliferation, accelerating apoptosis promotion and enhancing sensitization of XB1702 cells.@*Conclusions@#Silencing long-chain non-coding RNA TUG1 can enhance the radiosensitivity of cervical cancer cells. The mechanism may be related to targeting miR-145, which will provide a target for radiotherapy of cervical cancer.

Long Non-Coding RNA TUG1 Promotes Proliferation and Inhibits Apoptosis of Osteosarcoma Cells by Sponging miR-132-3p and Upregulating SOX4 Expression

PURPOSE: Long non-coding RNA taurine upregulated gene 1 (TUG1) is reported to be a vital regulator of the progression of various cancers. This study aimed to explore the exact roles and molecular mechanisms of TUG1 in osteosarcoma (OS) development. MATERIALS AND METHODS: Real-time quantitative PCR was applied to detect the expressions of TUG1 and microRNA-132-3p (miR-132-3p) in OS tissues and cells. Western blot was performed to measure protein levels of sex determining region Y-box 4 (SOX4). Cell viability was assessed using XTT assay. Cell apoptosis was evaluated using flow cytometry and caspase-3 activity detection assays. Bioinformatics analysis and luciferase reporter experiments were employed to confirm relationships among TUG1, miR-132-3p, and SOX4. RESULTS: TUG1 was highly expressed in human OS tissues, OS cell lines, and primary OS cells. TUG1 knockdown hindered proliferation and induced apoptosis in human OS cell lines and primary OS cells. Moreover, TUG1 inhibited miR-132-3p expression by direct interaction, and introduction of miR-132-3p inhibitor partly abrogated the effect of TUG1 knockdown on the proliferation and apoptosis of OS cells. Furthermore, SOX4 was validated as a target of miR-132-3p. Further functional analyses revealed that miR-132-3p inhibited proliferation and induced apoptosis of OS cells, while this effect was greatly abated following SOX4 overexpression. Moreover, TUG1 knockdown suppressed proliferation and promoted apoptosis by upregulating miR-132-3p and downregulating SOX4 in primary OS cells. CONCLUSION: TUG1 facilitated proliferation and suppressed apoptosis by regulating the miR-132-3p/SOX4 axis in human OS cell lines and primary OS cells. This finding provides a potential target for OS therapy.

Function of lncRNA TUG1 in Insulin Secretion from Pancreatic Beta Cells / 现代生物医学进展

Objective:To determine the role of lncRNA TUG1 in pancreatic β cells functioning both in vitro and in vivo.Methods:The lncRNA TUG1 expression in mice pancreas,brain,muscle and other different tissues was examined through qRT-PCR.MTT,flow cytometry,GSIS,ELISA and immunochemistry analyses were performed to detect the effect of lncRNA TUG1 on insulin secretion in vitro and in vivo.Results:lncRNA TUG1 was highly expressed in pancreatic tissue compared with other organ tissues.Knockdown of lncRNA TUG1 expression resulted in decreased insulin secretion in β cells both in vitro and in vivo.Immunochemistry analyses showed decreased relative islet area after treatment with lncRNA TUG1 siRNA.Conclusions:Downregulation of lncRNA TUG1 expression can affect insulin secretion in pancreatic β cells in vitro and in vivo,and lncRNA TUG1 may represent a factor that regulates the function of pancreatic β cells.

Expression of Long Non-coding RNA TUG1 and its Effect on Prognosis of Patients With Gastric Cancer / 胃肠病学

Background:The incidence of gastric cancer is gradually rising in recent years,long non-coding RNA taurine up-regulated gene 1 (TUG1)may have certain effects on the occurrence and progression of gastric cancer. Aims:To study the expression TUG1 in gastric cancer tissue and its effect on prognosis of patients with gastric cancer,and study the correlation between TUG1 and p27,cyclin D1. Methods:Surgically resected gastric cancer tissues and corresponding distal normal tissues of 48 gastric cancer patients from June 2013 to December 2013 at Qingdao Municipal Hospital were collected. qRT-PCR was used to detect the mRNA expression of TUG1,and its relationship with clinicopathological features was analyzed. Protein expressions of p27 and cyclin D1 were determined by Western blotting,and correlation with expression of TUG1 was analyzed. Kaplan-Meier was used to analyze the relationship between expression of TUG1 and prognosis. Results:The mRNA expression of TUG1 in gastric cancer tissues was significantly higher than that in corresponding normal tissues (6. 18 ± 0. 19 vs. 5. 09 ± 0. 16,P < 0. 05),and was not correlated with gender,age,tumor size,but correlated with lymph node metastasis,tumor differentiation and TNM staging (P < 0. 01). The protein expression of p27 was significantly decreased in gastric cancer tissues than in normal tissues (0. 1709 ± 0. 0212 vs. 0. 3087 ± 0. 0252,P < 0. 01),while protein expression of cyclin D1 was significantly increased (0. 3417 ± 0. 0271 vs. 0. 2417 ± 0. 0173,P < 0. 01),and the expression of p27 was negatively correlated with expression of cyclin D1 in gastric cancer tissues (r = - 0. 897,P < 0. 01). The expression of TUG1 was negatively correlated with expression of p27 (r = - 0. 730,P < 0. 01),and was positively correlated with expression of cyclin D1 (r = 0. 809,P < 0. 01)in gastric cancer tissues. The median survival time was shorter in gastric cancer patients with high-expressed TUG1 than in patients with low-expressed TUG1 (P < 0. 05). Conclusions:Long non-coding RNA TUG1 plays a role of cancer gene in the development of gastric cancer through p27 /cyclin D1 pathway. Detection of expression of TUG1 has important significance on the prediction of prognosis of gastric cancer patients.

Expression of Long Non-coding RNA TUG1 and its Effect on Prognosis of Patients With Gastric Cancer / 胃肠病学

Background:The incidence of gastric cancer is gradually rising in recent years,long non-coding RNA taurine up-regulated gene 1 (TUG1)may have certain effects on the occurrence and progression of gastric cancer. Aims:To study the expression TUG1 in gastric cancer tissue and its effect on prognosis of patients with gastric cancer,and study the correlation between TUG1 and p27,cyclin D1. Methods:Surgically resected gastric cancer tissues and corresponding distal normal tissues of 48 gastric cancer patients from June 2013 to December 2013 at Qingdao Municipal Hospital were collected. qRT-PCR was used to detect the mRNA expression of TUG1,and its relationship with clinicopathological features was analyzed. Protein expressions of p27 and cyclin D1 were determined by Western blotting,and correlation with expression of TUG1 was analyzed. Kaplan-Meier was used to analyze the relationship between expression of TUG1 and prognosis. Results:The mRNA expression of TUG1 in gastric cancer tissues was significantly higher than that in corresponding normal tissues (6. 18 ± 0. 19 vs. 5. 09 ± 0. 16,P < 0. 05),and was not correlated with gender,age,tumor size,but correlated with lymph node metastasis,tumor differentiation and TNM staging (P < 0. 01). The protein expression of p27 was significantly decreased in gastric cancer tissues than in normal tissues (0. 1709 ± 0. 0212 vs. 0. 3087 ± 0. 0252,P < 0. 01),while protein expression of cyclin D1 was significantly increased (0. 3417 ± 0. 0271 vs. 0. 2417 ± 0. 0173,P < 0. 01),and the expression of p27 was negatively correlated with expression of cyclin D1 in gastric cancer tissues (r = - 0. 897,P < 0. 01). The expression of TUG1 was negatively correlated with expression of p27 (r = - 0. 730,P < 0. 01),and was positively correlated with expression of cyclin D1 (r = 0. 809,P < 0. 01)in gastric cancer tissues. The median survival time was shorter in gastric cancer patients with high-expressed TUG1 than in patients with low-expressed TUG1 (P < 0. 05). Conclusions:Long non-coding RNA TUG1 plays a role of cancer gene in the development of gastric cancer through p27 /cyclin D1 pathway. Detection of expression of TUG1 has important significance on the prediction of prognosis of gastric cancer patients.

Inhibition of long non-coding RNA TUG1 on gastric cancer cell transference and invasion through regulating and controlling the expression of miR-144/c-Met axis

Objective To discuss the expression of long noncoding RNA TUG1 (lncRNA-TUG1) in gastric carcinoma (GC) and its effects on the transferring and invading capacity of gastric carcinoma cells. Methods Forty cases of carcinoma tissue and para-carcinoma tissue were selected from GC patients who underwent surgical removal in Zhejiang Provincial Hospital of Chinese Traditional Medicine and Wenzhou Central Hospital from January, 2013 to December, 2014; the expressing level of lncRNA-TUG1 in GC and para-C tissues was detected by applying the qRT-PCR technique. The correlation between lncRNA-TUG1 expression and patients' clinical data was classified and analyzed. SGC-7901 cells were transfected using lncRNA-TUG1 specific siRNA. Changes of the transferring and invading capacity of siRNA-transfected SGC-7901 cells were scratch-tested and transwell-detected. qRT-PCR was applied to detect the expression level of microRNA-144 after lncRNA-TUG1 was silenced. Changes of c-Met mRNA and protein expressions was detected by qRT-PCR and western-blot test. Results The expression level of lncRNA-TUG1 in GC tissue was significant higher than that in para-C tissue (P < 0.05) and the high expression level of lncRNA-TUG1 in GC tissue was significantly correlated with tumor lymph nodes metastasis and advance TNM phasing (P < 0.05). The transferring and invading capacity of SGC-7901 cells was highly inhibited after being transfected by lncRNA-TUG1 specific siRNA (P < 0.05). The results of qRT-PCR and western-blot proved that the expression of microRNA-144 was significantly boosted and the expression level of c-Met mRNA and protein was inhibited after lncRNA-TUG1 was silenced (P < 0.05). Conclusions lncRNA-TUG1 shows an up-regulated expression in GC tissue and that bears a correlation with clinicopathological features of malignant tumor. lncRNA-TUG1 can promote the transferring and invading capacity of GC by inhibiting the pathway of microRNA-144/c-Met.

Inhibition of long non-coding RNA TUG1 on gastric cancer cell transference and invasion through regulating and controlling the expression of miR-144/c-Met axis

OBJECTIVE@#To discuss the expression of long noncoding RNA TUG1 (lncRNA-TUG1) in gastric carcinoma (GC) and its effects on the transferring and invading capacity of gastric carcinoma cells.@*METHODS@#Forty cases of carcinoma tissue and para-carcinoma tissue were selected from GC patients who underwent surgical removal in Zhejiang Provincial Hospital of Chinese Traditional Medicine and Wenzhou Central Hospital from January, 2013 to December, 2014; the expressing level of lncRNA-TUG1 in GC and para-C tissues was detected by applying the qRT-PCR technique. The correlation between lncRNA-TUG1 expression and patients' clinical data was classified and analyzed. SGC-7901 cells were transfected using lncRNA-TUG1 specific siRNA. Changes of the transferring and invading capacity of siRNA-transfected SGC-7901 cells were scratch-tested and transwell-detected. qRT-PCR was applied to detect the expression level of microRNA-144 after lncRNA-TUG1 was silenced. Changes of c-Met mRNA and protein expressions was detected by qRT-PCR and western-blot test.@*RESULTS@#The expression level of lncRNA-TUG1 in GC tissue was significant higher than that in para-C tissue (P < 0.05) and the high expression level of lncRNA-TUG1 in GC tissue was significantly correlated with tumor lymph nodes metastasis and advance TNM phasing (P < 0.05). The transferring and invading capacity of SGC-7901 cells was highly inhibited after being transfected by lncRNA-TUG1 specific siRNA (P < 0.05). The results of qRT-PCR and western-blot proved that the expression of microRNA-144 was significantly boosted and the expression level of c-Met mRNA and protein was inhibited after lncRNA-TUG1 was silenced (P < 0.05).@*CONCLUSIONS@#lncRNA-TUG1 shows an up-regulated expression in GC tissue and that bears a correlation with clinicopathological features of malignant tumor. lncRNA-TUG1 can promote the transferring and invading capacity of GC by inhibiting the pathway of microRNA-144/c-Met.

LncRNA Taurine-Upregulated Gene 1 Promotes Cell Proliferation by Inhibiting MicroRNA-9 in MCF-7 Cells / 한국유방암학회지

PURPOSE: This study was designed to investigate the role of taurine-upregulated gene 1 (TUG1) in MCF-7 breast cancer cells and the molecular mechanism involved in the regulation of microRNA-9 (miR-9). METHODS: The expression of TUG1 in breast cancer tissues and cells was evaluated using quantitative reverse transcription polymerase chain reaction. Cell viability was examined using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay; cell cycle progression and apoptosis were analyzed using flow cytometry. A dual luciferase reporter assay was used to detect the relationship between TUG1 and miR-9. The expression of methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) was measured by western blot. RESULTS: Higher expression of TUG1 was observed in breast cancer tissues and cell lines than in the corresponding controls. TUG1 knockdown reduced proliferation, suppressed cell cycle progression, and promoted apoptosis of MCF-7 cells. The dual luciferase reporter assay showed that TUG1 could negatively regulate the expression of miR-9. MiR-9 inhibition abrogated the effect of TUG1 knockdown on the proliferation, cell cycle progression, and apoptosis of MCF-7 cells. TUG1 positively regulated the expression of MTHFD2 in breast cancer cells. CONCLUSION: TUG1 knockdown was significantly associated with decreased cell proliferation and it promoted apoptosis of breast cancer cells through the regulation of miR-9.