CRISPR-Cas13-Mediated Knockdown of lncRNA-GACAT3 Inhibited Cell Proliferation and Motility, and Induced Apoptosis by Increasing p21, Bax, and E-Cadherin Expression in Bladder Cancer

The current study is to investigate the expression pattern and biological function of long non-coding RNA Focally gastric cancer-associated transcript3 (GACAT3) in bladder cancer. Real-time quantitative qPCR was used to detect the expression level of GACAT-3 in tumor tissues and paired normal tissues. Human bladder cancer T24 and 5637 cell lines were transiently transfected with specific CRISPR-Cas13 or negative control CRISPR-Cas13. Cell migration, proliferation, and apoptosis were measured by using wound healing assay CCK-8 assay and Caspase-3 ELISA assay, respectively. The expression changes of p21, Bax, and E-cadherin after knockdown of GACAT3 were detected by using Western blot. The results demonstrated that GACAT3 was up-regulated in bladder cancer tissues than that in the paired normal tissues. Inhibition of cell proliferation, increased apoptosis, and decreased motility were observed in T24 and 5637 cell lines transfected by CRISPR-Cas13 targeting GACAT3. Downregulation of GACAT3 increased p21, Bax, and E-cadherin expression and silencing these genes could eliminate the phenotypic changes induced by knockdown of GACAT3. A ceRNA mechanism for GACAT3 was also revealed. By using CRISPR-Cas13 biotechnology, we suggested that GACAT3 may be a novel target for diagnosis and treatment of bladder cancer.

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Down-regulation LncRNA-SNHG15 contributes to proliferation and invasion of bladder cancer cells

BMC Urology ◽

10.1186/s12894-021-00852-1 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Aldhabi Mokhtar ◽

Chuize Kong ◽

Zhe Zhang ◽

Yan Du

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Tumor Stage ◽

Rt Pcr ◽

Down Regulation ◽

Bladder Cancer Cells ◽

Cancer Tissues ◽

Proliferation And Invasion

Abstract Objectives The aim of this study was to investigate the effect of lncRNA-SNHG15 in bladder carcinoma using cell lines experiments and the relationship between clinical characteristics and lncRNA-SNHG15 expression was analyzed. Methods Bladder cancer tissues and near-cancer tissues were collected. The real-time PCR (RT-PCR) was used to detect the expression of lncRNA-SNHG15 in tissues and cell lines. The expression of lncRNA-SNHG15 was downregulated by interference (siRNA), as detected by RT-PCR, that was used to determine the efficiency of the interference. CCK-8 and Transwell assays were used to evaluate the effect of lncRNA-SNHG15 on the proliferation and invasion capability of bladder cancer cells. The t-test was used for Statistical analyses, which were carried out using the Statistical Graph pad 8.0.1.224 software. Result The expression of lncRNA-SNHG15 was up regulated in 5637, UMUC3 and T24 cell lines compared with corresponding normal controls (P < 0.05). Up regulation was positively related to tumor stage (P = 0.015). And tumor size (P = 0.0465). The down-regulation of lncRNA-SNHG15 with siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion. Conclusion This study showed that lncRNA-SNHG15 is overexpressed in bladder cancer tissues and (5637, UMUC3 T24) cell lines. Up regulation was positively related to tumor stage (P = 0.015), and tumor size (P = 0.0465). Down-regulation of lncRNA-SNHG15 by siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion, indicating a potential molecular target for future tumor targeted therapy.

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ZFAS1 functions as an oncogenic long non-coding RNA in bladder cancer

Bioscience Reports ◽

10.1042/bsr20180475 ◽

2018 ◽

Vol 38 (3) ◽

Cited By ~ 10

Author(s):

Haifan Yang ◽

Ge Li ◽

Bo Cheng ◽

Rui Jiang

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Cancer Cell ◽

Biological Function ◽

Bladder Cancer Cell ◽

Migration And Invasion ◽

Cancer Cell Proliferation ◽

Non Coding Rna ◽

Cancer Tissues ◽

Long Non Coding Rna

Long non-coding RNA (lncRNA) ZFAS1 (zinc finger antisense 1) has been suggested to have an oncogenic role in the tumorigenesis of human malignant tumors. However, the expression status and biological function of ZFAS1 in bladder cancer is still unknown. Thus, the purpose of the present study is to explore the clinical value of ZFAS1 in bladder cancer patients, and the biological function of ZFAS1 in bladder cancer cell. In the present study, we found ZFAS1 expression was increased in bladder cancer tissues compared with paired adjacent normal tissues through analyzing the Cancer Genome Atlas (TCGA) database. Furthermore, we confirmed that levels of ZFAS1 expression were elevated in bladder cancer tissues and cell lines compared with normal bladder tissues and normal uroepithelium cell line, respectively. Then, we observed that the expression level of ZFAS1 was positively associated with clinical stag, muscularis invasion, lymph node metastasis, and distant metastasis in bladder cancer patients. The experiments in vitro suggested that knockdown of ZFAS1 repressed bladder cancer cell proliferation via up-regulating KLF2 and NKD2 expression, and inhibited cell migration and invasion via down-regulating ZEB1 and ZEB2 expression. In conclusion, ZFAS1 is overexpressed in bladder cancer, and functions as an oncogenic lncRNA in regulating bladder cancer cell proliferation, migration, and invasion.

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Expression and Biological Functions of EPC1 in Nasopharyngeal Carcinoma

Archives of Iranian Medicine ◽

10.34172/aim.2021.125 ◽

2021 ◽

Vol 24 (11) ◽

pp. 845-851

Author(s):

Yongmei Dai ◽

Wenhan Chen ◽

Chen Huang ◽

Shiyin Luo ◽

Junpeng Huang ◽

...

Keyword(s):

Nasopharyngeal Carcinoma ◽

Cell Lines ◽

Epithelial Mesenchymal Transition ◽

Malignant Tumors ◽

Vimentin Expression ◽

Mesenchymal Transition ◽

Induced Apoptosis ◽

And Function ◽

Cancer Tissues ◽

E Cadherin

Background: Comb homolog enhancer 1 (EPC1) gene is one of the important members of epigenetic inhibitor PCG family. It shows carcinogenic potential in a variety of malignant tumors, but the expression and role of EPC1 in nasopharyngeal carcinoma are unclear. The aim of this study was to explore the expression and function of enhancer of polycomb homolog 1 (EPC1) in nasopharyngeal carcinoma (NPC). Methods: The differential expression of EPC1 in the cancer tissues and cell lines of NPC was examined by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). EPC1 expression, cell proliferation, and apoptosis were detected in NPC cell lines after EPC1 silencing, and the levels of the epithelial-mesenchymal transition (EMT)-related proteins E-cadherin and vimentin were detected in NPC cells after EPC1 silencing. The study was performed at Fujian Provincial Hospital, Fujian, China, from 2018 to 2019. Results: We found that EPC1 was significantly upregulated in the cancer tissues and cell lines of NPC (P<0.001). Furthermore, knockdown of EPC1 inhibited the growth and metastasis of NPC cells. E-cadherin and vimentin were detected in NPC cells after EPC1 was knocked out. It was confirmed that inhibition of EPC1 resulted in increased E-cadherin expression (P<0.001) and decreased vimentin expression (P<0.001), suggesting that inhibition of EPC1 could inhibit the EMT in NPC cells. Conclusion: EPC1 expression was upregulated in NPC tissues and cell lines. Knockout of EPC1 effectively inhibited the growth of NPC cells, induced apoptosis, and inhibited invasion and metastasis. Inhibition of EPC1 could inhibit the EMT in NPC cells. All of the above findings support the viewpoint that EPC1 plays a pro-cancer role in NPC.

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Long non-coding RNA MIAT promotes cervical cancer proliferation and migration

The Journal of Biochemistry ◽

10.1093/jb/mvaa037 ◽

2020 ◽

Vol 168 (2) ◽

pp. 183-190 ◽

Cited By ~ 3

Author(s):

Lei Zhang ◽

Shuxia Ge ◽

Bing Cao

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cell Lines ◽

Cell Model ◽

Signalling Pathway ◽

Migration And Invasion ◽

Non Coding Rna ◽

Cancer Tissues ◽

Long Non Coding Rna ◽

Proliferation And Invasion

Abstract Cervical cancer is one of the most common cancers in the world while its pathological mechanisms are not well-elucidated. Long non-coding RNA (lncRNA) has been implicated in cancer development. The dysregulation of lncRNA myocardial infarction-associated transcript (MIAT) has been reported in several cancers while its role in cervical cancer is not described yet. In this study, the role of MIAT in cervical cancer was explored. We evaluated the expression of MIAT in cervical cancer tissues and cell lines. Furthermore, we explored the effects of MIAT on proliferation and invasion of cervical cancer using cell model and animal transplantation model. We also evaluated the effects of MIAT on activation of PI3K/Akt/mTOR signalling pathway. Our results show that MIAT was up-regulated in cervical cancer tissues and cell lines. Knocking down MIAT resulted in decreased cell proliferation, migration and invasion of cervical cancer cells and suppression of tumour growth in mice. Mechanically, knocking down MIAT suppressed the activation of PI3K/Akt signalling pathway. In conclusion, MIAT promotes cell proliferation and invasion in cervical cancer.

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Cell proliferation in kidney carcinoma is inhibited by lncRNA GASL1

European Journal of Inflammation ◽

10.1177/2058739219854598 ◽

2019 ◽

Vol 17 ◽

pp. 205873921985459

Author(s):

Jianping Dong ◽

Shiping Zheng ◽

Xiaoyan Yang ◽

Xiuyan Song

Keyword(s):

Cell Proliferation ◽

Cell Line ◽

Cell Lines ◽

Cancer Cell ◽

Kidney Cancer ◽

Noncoding Rna ◽

Cancer Cell Line ◽

Cancer Cell Lines ◽

Normal Tissues ◽

Cancer Tissues

Long noncoding RNA (lncRNA) GASL1 was identified as a novel lncRNA, which plays an important role in the proliferation and apoptosis of cells. This study aimed to compare the expression of GASL1 mRNA in kidney cancer cells and normal cells and detect the biological role of GASL1 in kidney cancer cell line A498. Polymerase chain reaction (PCR) was performed to examine the expression of GASL1 mRNA in kidney cancer tissues, normal tissues, and the cell lines. GASL1 overexpression was achieved in kidney cancer cell lines A498 through transfection. MTT was used to detect the effects of GASL1 overexpression in A498 cells. GASL1 mRNA was significantly overexpressed in adjacent normal tissues compared with renal cell carcinoma. The expression of GASL1 is lower in kidney cancer cell lines than in normal kidney epithelium cell line HREpiC. Overexpression of GASL1 inhibits the proliferation of renal carcinoma cell lines. GASL1 mRNA was down-regulated in kidney cancer tissues and may play a role in kidney cancer cell proliferation.

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Knockdown of TPT1-AS1 inhibits cell proliferation, cell cycle G1/S transition, and epithelial–mesenchymal transition in gastric cancer

Bosnian Journal of Basic Medical Sciences ◽

10.17305/bjbms.2020.4470 ◽

2020 ◽

Cited By ~ 3

Author(s):

Jun Tang ◽

Fei Huang ◽

Hui Wang ◽

Feng Cheng ◽

Yaping Pi ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Cycle ◽

Cell Proliferation ◽

Cell Lines ◽

Cell Cycle Progression ◽

Epithelial Mesenchymal Transition ◽

Negative Control ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

Normal Tissues

Long non-coding RNAs are considered to be critical regulators of tumor progression. Tumor protein translationally controlled 1 antisense RNA 1 (TPT1-AS1) was shown to have an oncogenic role in cervical and ovarian cancer. The clinical significance and biological function of TPT1-AS1 in gastric cancer (GC) are not clear. In this study, we analyzed the expression of TPT1-AS1 in GC tissues and cell lines and performed functional and mechanistic analysis of TPT1-AS1 effects on GC cell proliferation, migration, and invasion. TPT1-AS1 expression was determined in 76 pairs of GC tissues vs. matched adjacent normal tissues and in four GC cell lines (SGC-7901, AGS, BGC-823, and MGC-803) vs. GES-1 cell line by quantitative reverse transcription PCR. SGC-7901 and MGC-803 cells were transfected with small interfering RNA or scrambled negative control, and cell proliferation, colony formation, migration, invasion and cell cycle assays were performed. The expression of proteins involved in cell cycle progression and epithelial–mesenchymal transition was analyzed by Western blot. TPT1-AS1 expression was significantly higher in GC tissues and cell lines compared to controls. The overexpression of TPT1-AS1 was significantly correlated with TNM stage and lymph node metastasis, and it was associated with worse prognosis of GC patients according to the Kaplan–Meier survival analysis and Cox proportional hazard regression analysis. The knockdown of TPT1-AS1 significantly inhibited proliferation, cell cycle G1/S transition, migration, and invasion of SGC-7901 and MGC-803 cells. Moreover, TPT1-AS1 knockdown downregulated the expression of CDK4, cyclin D1, and vimentin and upregulated the expression of p21 and E-cadherin. Our findings suggest that TPT1-AS1 may be a promising therapeutic target in GC.

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Up-Regulating ERIC by CRISPR-dCas9-VPR Inhibits Cell Proliferation and Invasion and Promotes Apoptosis in Human Bladder Cancer

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.654718 ◽

2021 ◽

Vol 8 ◽

Author(s):

Jiangeng Yang ◽

An Xia ◽

Huajie Zhang ◽

Qi Liu ◽

Hongke You ◽

...

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Urothelial Carcinoma ◽

Cell Lines ◽

Expression Patterns ◽

Regulation Of Gene Expression ◽

Negative Control Group ◽

Negative Control ◽

Control Group ◽

Bladder Urothelial Carcinoma

LncRNAs are defined as non-coding RNAs that are longer than 200 nucleotides in length. The previous studys has shown that lncRNAs played important roles in the regulation of gene expression and were essential in mammalian development and disease processes. Inspired by the observation that lncRNAs are aberrantly expressed in tumors, we extracted RNA from Bladder urothelial carcinoma and matched histologically normal urothelium from each patient and bladder carcinoma cell lines. Then, we reversed transcribed them into cDNA.Last, we investigated the expression patterns of ERIC by the fluorescence quantitative PCR in bladder cancer tissues and cell lines. CRISPR-dCas9-VPR targeting ERIC plasmid was transfected into T24 and 5637 cells, and cells were classified into two groups: negative control (NC) and ERIC overexpression group. MTT assay, transwell assay, and flow cytometry were performed to examine changes in cell proliferation, invasiveness, and apoptosis. We found that the expression of ERIC was down-regulated in bladder urothelial carcinoma compared to matched histologically normal urotheliam. The differences of the expression of this gene were large in the bladder cancer lines. Compared with the negative control group, the ERIC overexpression group showed significantly decreased cell proliferation rate (t = 7.583, p = 0.002; t = 3.283, p = 0.03) and invasiveness (t = 11.538, p < 0.001; t = 8.205, p = 0.01); and increased apoptotic rate (t = −34.083, p < 0.001; t = −14.316, p < 0.001). Our study lays a foundation for further study of its pathogenic mechanism in bladder cancer.

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CRISPR-CasRx Targeting LncRNA LINC00341 Inhibits Tumor Cell Growth in vitro and in vivo

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.638995 ◽

2021 ◽

Vol 8 ◽

Author(s):

Chunjing Li ◽

Yu Cao ◽

Li Zhang ◽

Jierong Li ◽

Jianfeng Wang ◽

...

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Cell Lines ◽

Low Grade ◽

Human Bladder Cancer ◽

Human Bladder ◽

Tumor Tissues ◽

E Cadherin

CRISPR-CasRx technology provides a new and powerful method for studying cellular RNA in human cancer. Herein, the pattern of expression of long noncoding RNA 00341 (LINC00341) as well as its biological function in bladder cancer were studied using CRISPR-CasRx. qRT-PCR was employed to quantify the levels of expression of LINC00341 in tumor tissues along with the matched non-tumor tissues. sgRNA targeting LINC00341 or the sgRNA negative control were transiently transfected into the T24 as well as 5,637 human bladder cancer cell lines. CCK-8, ELISA as well as wound healing methods were employed to explore cell proliferation, apoptosis and migration, respectively. The tumorigenicity experiment in nude mice also performed to detect cell proliferation. The expression of p21, Bax as well as E-cadherin were assayed using western blot. The results demonstrated that LINC00341 was overexpressed in bladder cancer in contrast with the healthy tissues. The LINC00341 expression level in high-grade tumors was higher in contrast with that in low-grade tumors. The expression of linc00341 was higher relative to that of non-invasive tumors. In T24 as well as 5637-cell lines harboring LINC00341-sgRNA, inhibition of cell proliferation (in vitro and in vivo), elevated apoptosis rate and diminished migration ability. Moreover, silencing LINC00341 upregulated the expressions of p21, Bax as well as E-cadherin. Knockout of these genes could eliminate the phenotypic changes caused by sgRNA targeting LINC00341. Our data demonstrate that LINC00341 has a carcinogenic role in human bladder cancer.

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Down-regulation LncRNA-SNHG15 Contributes to Proliferation and Invasion of Bladder Cancer Cells

10.21203/rs.3.rs-179702/v1 ◽

2021 ◽

Author(s):

Aldhabi Mokhtar ◽

Chuize Kong ◽

Zhe Zhang ◽

Yan Du

Keyword(s):

Cell Proliferation ◽

Bladder Cancer ◽

Cell Lines ◽

Tumor Size ◽

Tumor Stage ◽

Rt Pcr ◽

Down Regulation ◽

Statistical Graph ◽

Cancer Tissues ◽

Proliferation And Invasion

Abstract OBJECTIVES: This study aimed to investigate effect of lncRNA-SNHG15 in bladder carcinoma using cell lines experiments and the relationship between clinical characteristics and lncRNA-SNHG15 expression was analyzed.Methods. Bladder cancer tissues and near-cancer tissues were collected. The expression of lncRNA-SNHG15 in tissues and cell lines was detected by real-time PCR (RT-PCR). The expression of lncRNA-SNHG15 was downregulated by interference (siRNA) as detected by RT-PCR that was used to detect the interference efficiency. CCK-8, and Transwell assays were used to evaluate the effect of lncRNA-SNHG15 on the proliferation, invasion capability of bladder cancer cells.t-test was used for Statistical analyses, were performed using the Statistical Graph pad 8.0.1.224 software.Result: The expression of lncRNA-SNHG15 was up regulated in 5637, UMUC3 and T24 cell lines compared with corresponding normal controls (P<0.05). up regulation was positively related to tumor stage (P = 0.015), and tumor size (P =0.0465) . The down-regulation of lncRNA-SNHG15 with siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion.Conclusion: This study showed that lncRNA-SNHG15 is overexpressed in bladder cancer tissues and (5637, UMUC3 T24) cell lines. up regulation was positively related to tumor stage (P = 0.015), and tumor size (P =0.0465) .The down-regulation of lncRNA-SNHG15 with siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion, which provides a potential molecular target for future tumor targeted therapy.

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Long Non-coding RNA PANDAR Promotes Radioresistance in Nasopharyngeal Carcinoma via SIRT1/Ku70/PI3K/Akt Pathway

10.21203/rs.3.rs-762963/v1 ◽

2021 ◽

Author(s):

Si-wei Li ◽

Guo-liang Pi ◽

Yong Zeng ◽

Chang-li Ruan ◽

Xiao-song He ◽

...

Keyword(s):

Cell Proliferation ◽

Dna Damage ◽

Nasopharyngeal Carcinoma ◽

Sirtuin 1 ◽

Akt Pathway ◽

Phosphatidylinositol 3 Kinase ◽

Normal Tissues ◽

Non Coding Rna ◽

Cancer Tissues ◽

Long Non Coding Rna

Abstract Objective: Radioresistance may result in nasopharyngeal carcinoma (NPC) radiation failure. To comprehend the concrete mechanism in NPC radioresistance, this work was initiated from long non-coding RNA (lncRNA) promoter of CDKN1A antisense DNA damage activated RNA (PANDAR), accompanied by sirtuin 1 (SIRT1), Ku70, and phosphatidylinositol 3 kinase (PI3K)/Akt pathway.Methods: NPC cancer tissues and normal tissues were harvested and NPC cancer tissues were specified into radioresistant and radiosensitive types. The connection between PANDA expression with NPC radioresistance, clinicopathological traits and prognosis was tested. Radioresistant CNE2-IR and 5-8F-IR cells were induced and transfected with depleted PANDAR or SIRT1 to identify their roles in cell proliferation, cycle distribution, apoptosis, SIRT1, Ku70 and PI3K/Akt pathway. PANDA and SIRT1 expression in CNE2-IR and 5-8F-IR cells were tested.Results: PANDA was elevated in NPC tissues and radioresistant tissues relative to normal tissues and radiosensitive tissues. Raised PANDA was connected with NPC radioresistance and unsatisfactory prognosis. CNE2-IR and 5-8F-IR cells expressed up-regulated PANDA and SIRT1. Down-regulating PANDA or SIRT1 inhibited radioresistant NPC cell proliferation, decreased SIRT1 and Ku70, and inactivated PI3K/Akt pathway.Conclusion: This work has clued that depleting PANDA decreases SIRT1 recruitment to inactivate Ku70 deacetylation-mediated PI3K/Akt pathway, thereby promoting NPC radiosensitivity.

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