Lipid Coated and Chlorin e6 Loaded Calcium Carbonate for Effective In Situ Immunotheraphy of Colorectal Cancer

2020 ◽  
Vol 16 (8) ◽  
pp. 1196-1204
Author(s):  
Wenxin Dai ◽  
Shaoquan Lu ◽  
Wangyuan Zeng ◽  
Dongwon Lee
Keyword(s):  
Colorectal Cancer ◽  
Calcium Carbonate ◽  
Cancer Vaccine ◽  
Inflammatory Responses ◽  
Chlorin E6 ◽  
Strong Immune Response ◽  
Tumor Associated Antigen

Cancer vaccine is well recognized as a novel but effective way for cancer immunotherapy. Especially, the role of dendritic cells (DCs) in antigen presentation properties is critical for the final performance of cancer vaccine. Herein, a lipid (Li) coated calcium carbonate (CC) vehicle (Li/CC) was employed to load chlorin e6 (Ce6) to serve as a potential in situ vaccine (Li/CC-Ce6) for effective immunotherapy of colorectal cancer. It was suggested that the loaded Ce6 within Li/CCCe6 can be activated under laser irradiation. The photodynamic therapy (PDT) of Ce6 was expected to produce reactive oxygen species (ROS) to cause cell death and expose tumor-associated antigen (TAA). In addition, the produced ROS can mimic the inflammatory responses for the recruitment of DC to initiate strong immune response cascade. Moreover, the recruitment of DC can recognize the exposed TAA to stimulate DC for effective vaccination in situ. Results from in vitro and in vivo assays demonstrated the strong ability of this platform to enhance DC vaccination, resulting in promising growth inhibition of both primary and distant tumors.

scholarly journals hsa_circ_0000231 Promotes Colorectal Cancer Cell Growth Through Upregulation of CCND2 by IGF2BP3/miR-375 Dual Pathway

2020 ◽  
Author(s):  
Wei Zhang ◽  
Bo Wang ◽  
Yang Yang ◽  
Zhen Zhang ◽  
Quan Wang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
In Situ Hybridization ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Cancer Cell Growth ◽  
Normal Tissues

Abstract Background circular RNAs (circRNAs) recently have been emerged as vital regulators for involvement of initiation and progression of diverse kinds of human cancers. This study aimed to investigate the role of circRNAs in colorectal cancer (CRC). Methods The expression profile of circRNAs in 5 pairs of CRC tissues and adjacent normal tissues were analyzed by Microarray. Quantitative real-time PCR and in situ hybridization and Base Scope Assay were used to determine the level and prognostic values of hsa_circ_0000231. Then, functional experiments in vitro and in vivo were performed to investigate the effects of hsa_circ_0000231 on cell proliferation. Mechanistically, fluorescent in situ hybridization, dual luciferase reporter assay, RNA pull-down and RNA immunoprecipitation experiments were performed to confirm the interaction between hsa_circ_0000231 and IGF2BP3 or has_miR-375. Results hsa_circ_0000231 was evidently up-regulated in CRC primary tissues, which was indicated to poor prognosis of CRC patients. The results demonstrated that hsa_circ_0000231 could promote CRC cell proliferation as well as tumorigenesis in vitro and in vivo. Mechanistic analysis showed that hsa_circ_0000231 might on the one hand act as a ceRNA (competing endogenous RNA) of miR-375 to regulate cyclin D2 (CCND2), and on the other hand bind to IGF2BP3 protein to protect CCND2 from being degraded. Conclusion Our findings suggest that hsa_circ_0000231 facilitated CRC progression by sponging miR-375 or binding to IGF2BP3 to modulate CCND2. This discovery implied that has_circ_0000231 may be a potential new diagnostic and therapeutic biomarker for CRC.

scholarly journals Long noncoding RNA RP11-757G1.5 sponges miR-139-5p and upregulates YAP1 thereby promoting the proliferation and liver, spleen metastasis of colorectal cancer

2020 ◽  
Vol 39 (1) ◽  
Author(s):  
Xiaojian Zhu ◽  
Fanqin Bu ◽  
Ting Tan ◽  
Qilin Luo ◽  
Jinfeng Zhu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
In Situ Hybridization ◽  
Cox Regression ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Cox Regression Analysis

Abstract Background Accumulating evidence indicates that long non-coding RNAs (lncRNAs) acting as crucial regulators in tumorigenesis. However, its biological functions of lncRNAs in colorectal cancer (CRC) have not been systematically clarified. Methods An unbiased screening was performed to identify disregulated lncRNAs revealed to be implicated in CRC carcinogenesis according to an online-available data dataset. In situ hybridization (ISH), RT-qPCR and RNA fluorescence in situ hybridization (RNA-FISH) were applied to detect RP11-757G1.5 expression in CRC tissues and cell lines. The associations of RP11-757G1.5 with clinicopathological characteristics were analyzed. Their effects on prognosis were analyzed by the Kaplan-Meier analysis, Log-rank test, Univariate and Multivariate Cox regression analysis. The potential biological function of RP11-757G1.5 in CRC was investigated by Colony formation, Edu cell proliferation, Flow cytometry, Wound healing and Transwell assays. Bioinformatics binding site analysis, Luciferase reporter assay, Ago2 immunoprecipitation assays, RNA pull-down assay, RT-qPCR and Western blotting were utilized to demonstrate the mechanism of RP11-757G1.5 acts as a molecular sponge of miR-139-5p to regulate the expression of YAP1. Finally, we further explore the potential role of RP11-757G1.5 in CRC orthotopic xenografts in vivo. Results We discovered a novel oncogenic lncRNA RP11-757G1.5, that was overexpressed in CRC tissues, especially in aggressive cases. Moreover, up-regulation of RP11-757G1.5 strongly correlated with poor clinical outcomes of patients with CRC. Functional analyses revealed that RP11-757G1.5 promoted cell proliferation in vitro and in vivo. Furthermore, RP11-757G1.5 stimulated cell migration and invasion in vitro and in vivo. Mechanistic studies illustrated that RP11-757G1.5 regulated the expression of YAP1 through sponging miR-139-5p and inhibiting its activity thereby promoting CRC progression and development. Conclusions Altogether, these results reveal a novel RP11-757G1.5/miR-139-5p/YAP1 regulatory axis that participates in CRC carcinogenesis and progression.

scholarly journals Circ3823 contributes to growth, metastasis and angiogenesis of colorectal cancer: involvement of miR-30c-5p/TCF7 axis

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Yaxin Guo ◽  
Yuying Guo ◽  
Chen Chen ◽  
Dandan Fan ◽  
Xiaoke Wu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
In Situ Hybridization ◽  
Tumour Growth ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Repressive Effect ◽  
Underlying Mechanisms

Abstract Background Colorectal cancer (CRC) is one of the most common malignant tumours. The recurrence and metastasis of CRC seriously affect the survival rate of patients. Angiogenesis is an extremely important cause of tumour growth and metastasis. Circular RNAs (circRNAs) have been emerged as vital regulators for tumour progression. However, the regulatory role, clinical significance and underlying mechanisms still remain largely unknown. Methods High-throughput sequencing was used to analyse differential circRNAs expression in tumour and non-tumour tissues of CRC. In situ hybridization (ISH) and qRT-PCR were used to determine the level of circ3823 in CRC tissues and serum samples. Then, functional experiments in vitro and in vivo were performed to investigate the effects of circ3823 on tumour growth, metastasis and angiogenesis in CRC. Sanger sequencing, RNase R and Actinomycin D assay were used to verify the ring structure of circ3823. Mechanistically, dual luciferase reporter assay, fluorescent in situ hybridization (FISH), RNA immunoprecipitation (RIP) and RNA pull-down experiments were performed to confirm the underlying mechanisms of circ3823. Results Circ3823 was evidently highly expressed in CRC and high circ3823 expression predicted a worse prognosis of CRC patients. Receiver operating characteristic curves (ROCs) indicated that the expression of circ3823 in serum showed high sensitivity and specificity for detecting CRC which means circ3823 have the potential to be used as diagnostic biomarkers. Functional experiments in vitro and in vivo indicated that circ3823 promote CRC cell proliferation, metastasis and angiogenesis. Mechanism analysis showed that circ3823 act as a competing endogenous RNA of miR-30c-5p to relieve the repressive effect of miR-30c-5p on its target TCF7 which upregulates MYC and CCND1, and finally facilitates CRC progression. In addition, we found that N6-methyladenosine (m6A) modification exists on circ3823. And the m6A modification is involved in regulating the degradation of circ3823. Conclusions Our findings suggest that circ3823 promotes CRC growth, metastasis and angiogenesis through circ3823/miR-30c-5p/TCF7 axis and it may serve as a new diagnostic marker or target for treatment of CRC patients. In addition, m6A modification is involved in regulating the degradation of circ3823.

scholarly journals Long noncoding RNA RP11-757G1.5 sponges miR-139-5p and upregulates YAP1 thereby promoting the proliferation and liver, spleen metastasis of colorectal cancer

2020 ◽  
Author(s):  
xiaojian zhu ◽  
Fanqin Bu ◽  
Ting Tan ◽  
Qilin Luo ◽  
Jingfeng Zhu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
In Situ Hybridization ◽  
Cox Regression ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Cox Regression Analysis

Abstract Background: Accumulating evidence indicates that long non-coding RNAs (lncRNAs) acting as crucial regulators in tumorigenesis. However, its biological functions of lncRNAs in colorectal cancer (CRC) have not been systematically clarified. Methods: An unbiased screening was performed to identify disregulated lncRNAs revealed to be implicated in CRC carcinogenesis according to an online-available data dataset. In situ hybridization (ISH), RT-qPCR and RNA fluorescence in situ hybridization (RNA-FISH) were applied to detect RP11-757G1.5 expression in CRC tissues and cell lines. The associations of RP11-757G1.5 with clinicopathological characteristics were analyzed. Their effects on prognosis were analyzed by the Kaplan-Meier analysis, Log-rank test, Univariate and Multivariate Cox regression analysis. The potential biological function of RP11-757G1.5 in CRC was investigated by Colony formation, Edu cell proliferation, Flow cytometry, Wound healing and Transwell assays. Bioinformatics binding site analysis, Luciferase reporter assay, Ago2 immunoprecipitation assays, RNA pull-down assay, RT-qPCR and Western blotting were utilized to demonstrate the mechanism of RP11-757G1.5 acts as a molecular sponge of miR-139-5p to regulate the expression of YAP1. Finally, we further explore the potential role of RP11-757G1.5 in CRC orthotopic xenografts in vivio . Results: We discovered a novel oncogenic lncRNA RP11-757G1.5, that was overexpressed in CRC tissues, especially in aggressive cases. Moreover, up-regulation of RP11-757G1.5 strongly correlated with poor clinical outcomes of patients with CRC. Functional analyses revealed that RP11-757G1.5 promoted cell proliferation in vitro and in vivo . Furthermore, RP11-757G1.5 stimulated cell migration and invasion in vitro and in vivo . Mechanistic studies illustrated that RP11-757G1.5 regulated the expression of YAP1 through sponging miR-139-5p and inhibiting its activity thereby promoting CRC progression and development. Conclusions: Altogether, these results reveal a novel RP11-757G1.5/miR-139-5p/YAP1 regulatory axis that participates in CRC carcinogenesis and progression.

hsa_circ_0000231 promotes colorectal cancer cell growth through upregulation of CCND2 by IGF2BP3/miR-375 dual pathway 

2020 ◽  
Author(s):  
Wei Zhang ◽  
Bo Wang ◽  
Yang Yang ◽  
Zhen Zhang ◽  
Quan Wang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
Expression Profiles ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Cancer Cell Growth ◽  
Normal Tissues

Abstract Background and aim Circular RNAs (circRNAs) have emerged as vital regulators of the initiation and progression of diverse kinds of human cancers. This study aimed to investigate the role of circRNAs in colorectal cancer (CRC). Methods The expression profiles of circRNAs in five pairs of CRC tissues and adjacent normal tissues were analyzed using microarray. Quantitative real-time polymerase chain reaction, in situ hybridization, and BaseScope Assay were used to determine the level and prognostic values of hsa_circ_0000231. Then, in vitro and in vivo functional experiments were performed to investigate the effects of hsa_circ_0000231 on cell proliferation. Mechanistically, fluorescence in situ hybridization, dual-luciferase reporter assay, and RNA pull-down and RNA immunoprecipitation experiments were performed to confirm the interaction between hsa_circ_0000231 and Insulin-like growth factor 2 mRNA-binding protein 3(IGF2BP3) or has_miR-375. Results The expression of hsa_circ_0000231 was upregulated in CRC primary tissues, which indicated poor prognosis of patients with CRC. The results demonstrated that hsa_circ_0000231 could promote CRC cell proliferation as well as tumorigenesis in vitro and in vivo. The mechanistic analysis showed that hsa_circ_0000231 might, on the one hand, act as a competing endogenous RNA of miR-375 to promote cyclin D2 (CCND2) and, on the other hand, bind to the IGF2BP3 protein to prevent CCND2 degradation. Conclusion The findings suggested that hsa_circ_0000231 facilitated CRC progression by sponging miR-375 or binding to IGF2BP3 to modulate CCND2, implying that has_circ_0000231 might be a potential new diagnostic and therapeutic biomarker of CRC.

Development of floating in situ gelling system as an efficient anti-ulcer formulation: in vitro and in vivo studies

RSC Advances ◽  
2015 ◽  
Vol 5 (37) ◽  
pp. 28848-28856 ◽  
Author(s):  
C. Bothiraja ◽  
Vijay Kumbhar ◽  
Atmaram Pawar ◽  
Karimunnisa Shaikh ◽  
Ravindra Kamble
Keyword(s):  
Calcium Carbonate ◽  
Gellan Gum ◽  
In Vivo Studies ◽  
Model Drug ◽  
In Situ Gelling

The aim of the present work was to design gellan gum and calcium carbonate based floating in situ gel as an efficient anti-ulcer formulation using andrographolide (AG) as a model drug.

scholarly journals RNA methylation-mediated LINC01559 suppresses colorectal cancer progression by regulating the miR-106b-5p/PTEN axis

2021 ◽  
Author(s):  
Ke Shi ◽  
Shuaixi Yang ◽  
Quanbo Zhou ◽  
Chen Chen ◽  
Bo Shao ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
In Situ Hybridization ◽  
Noncoding Rnas ◽  
Bioinformatic Analysis ◽  
Migration And Invasion ◽  
Sequencing Analysis ◽  
Rna Methylation

Abstract BackgroundLong noncoding RNAs (lncRNAs) regulate multiple biological effects in cancers. Recently, RNA methylation has been found to modify not only coding RNAs but also some noncoding RNAs. How RNA methylation affects lncRNAs to affect colorectal cancer (CRC) progression remains elusive.MethodsRNA-sequencing of cancer and normal tissues was analysed. LINC01559 in CRC was selected and verified by western blot (WB), quantitative real-time PCR (qRT-PCR) and in situ hybridization (ISH). Functional experiments in vitro and in vivo were used to explore the biological functions of LINC01559 in CRC. The LINC01559/miR-106-5p/PTEN axis was explored at different levels of analysis, including fluorescence in situ hybridization (FISH), luciferase assays, and rescue experiments. RIP-sequencing, m6A RNA immunoprecipitation (MeRIP) assays and bioinformatic analysis were conducted to determine the upstream mechanism of LINC01559.ResultsLINC01559, identified by RNA-sequencing analysis, was downregulated in CRC tissues and cell lines compared with normal controls. Lower expression of LINC01559 in CRC patients predicted a poor prognosis. In addition, PTEN was found to be positively correlated with LINC01559, and bioinformatic analysis and in vitro experiments indicated that miR-106b-5p could be the link between LINC01559 and PTEN. Then, downregulated LINC01559 restored the malignant phenotype of CRC cells in vivo and in vitro, while cotransfection of si-LINC01559 and miR-106b-5p inhibitor neutralized this effect. Notably, METTL3 was found to be highly expressed in CRC, and the downregulation of METTL3 could inhibit the migration and invasion of CRC cells. Mechanistically, we found abundant m6A modification sites on LINC01559. Then, we uncovered these sites as potential targets of METTL3 through experiments in vivo.ConclusionThe results revealed a negative functional regulation of LINC01559/miR-106b-5p/PTEN axis in CRC progression, and explored a new mechanism of METTL3-mediated m6A modification on LINC01559 formation. These results elucidate a novel potential therapeutic target for CRC treatment.Trial registrationThe human cancer tissues used in this study were approved by Ethnics Committee of The First Affiliated Hospital of Zhengzhou University in December 19, 2019, and the TRN is 2019-KW-423.

NLRP3 Promotes Colorectal Cancer Cell Proliferation and Metastasis via Regulating Epithelial Mesenchymal Transformation

2020 ◽  
Vol 20 (7) ◽  
pp. 820-827 ◽  
Author(s):  
Xinyu Shao ◽  
Zhiyi Lei ◽  
Chunli Zhou
Keyword(s):  
Colorectal Cancer ◽  
Molecular Mechanisms ◽  
Epithelial Mesenchymal Transition ◽  
Inflammatory Responses ◽  
Venous Invasion ◽  
Tnm Staging ◽  
Mesenchymal Transition ◽  
Mesenchymal Transformation

Background: Nucleotide-binding domain Leucine-rich Repeat Protein 3 (NLRP3) plays a regulatory role in the immune and inflammatory responses, and has been implicated in Colorectal Cancer (CRC) progression and metastasis. However, the underlying molecular mechanisms have not been fully elucidated. Methods: In this study, we analyzed the expression levels of NLRP3 in human CRC tissues, and performed functional assays in CRC cell lines and a subcutaneous tumor model to elucidate its role in the development and progression of CRC. Results: In this study, we found that NLRP3 was significantly upregulated in human CRC tissues and was associated with tumor size and invasion, lymph node metastasis, venous invasion, neural invasion and TNM staging. Furthermore, knockdown of NLRP3 in CRC cells inhibited their migration and growth in vitro and in vivo, and reversed Epithelial-Mesenchymal Transition (EMT) in vitro. Conclusion: Our findings indicate that NLRP3 likely regulates CRC metastasis by activating the EMT program, and is a potential therapeutic target.

scholarly journals hsa_circ_0000231 promotes colorectal cancer cell growth through upregulation of CCND2 by IGF2BP3/miR-375 dual pathway

2020 ◽  
Author(s):  
Wei Zhang ◽  
Bo Wang ◽  
Yang Yang ◽  
Zhen Zhang ◽  
Quan Wang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
Expression Profiles ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Cancer Cell Growth ◽  
Normal Tissues

Abstract Background and aim Circular RNAs (circRNAs) have emerged as vital regulators of the initiation and progression of diverse kinds of human cancers. This study aimed to investigate the role of circRNAs in colorectal cancer (CRC).Methods The expression profiles of circRNAs in five pairs of CRC tissues and adjacent normal tissues were analyzed using microarray. Quantitative real-time polymerase chain reaction, in situ hybridization, and BaseScope Assay were used to determine the level and prognostic values of hsa_circ_0000231. Then, in vitro and in vivo functional experiments were performed to investigate the effects of hsa_circ_0000231 on cell proliferation. Mechanistically, fluorescence in situ hybridization, dual-luciferase reporter assay, and RNA pull-down and RNA immunoprecipitation experiments were performed to confirm the interaction between hsa_circ_0000231 and Insulin-like growth factor 2 mRNA-binding protein 3(IGF2BP3) or has_miR-375. Results The expression of hsa_circ_0000231 was upregulated in CRC primary tissues, which indicated poor prognosis of patients with CRC. The results demonstrated that hsa_circ_0000231 could promote CRC cell proliferation as well as tumorigenesis in vitro and in vivo. The mechanistic analysis showed that hsa_circ_0000231 might, on the one hand, act as a competing endogenous RNA of miR-375 to promote cyclin D2 (CCND2) and, on the other hand, bind to the IGF2BP3 protein to prevent CCND2 degradation. Conclusion The findings suggested that hsa_circ_0000231 facilitated CRC progression by sponging miR-375 or binding to IGF2BP3 to modulate CCND2, implying that has_circ_0000231 might be a potential new diagnostic and therapeutic biomarker of CRC.

scholarly journals Long noncoding RNA RP11-757G1.5 sponges miR-139-5p and upregulates YAP1 thereby promoting the proliferation and liver, spleen metastasis of colorectal cancer

2020 ◽  
Author(s):  
Xiaojian Zhu ◽  
Fanqin Bu ◽  
Ting Tan ◽  
Qilin Luo ◽  
Jingfeng Zhu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Proliferation ◽  
In Situ Hybridization ◽  
Cox Regression ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Cox Regression Analysis

Abstract Background: Accumulating evidence indicates that long non-coding RNAs (lncRNAs) acting as crucial regulators in tumorigenesis. However, its biological functions of lncRNAs in colorectal cancer (CRC) have not been systematically clarified.Methods: An unbiased screening was performed to identify disregulated lncRNAs revealed to be implicated in CRC carcinogenesis according to an online-available data dataset. In situ hybridization (ISH), RT-qPCR and RNA fluorescence in situ hybridization (RNA-FISH) were applied to detect RP11-757G1.5 expression in CRC tissues and cell lines. The associations of RP11-757G1.5 with clinicopathological characteristics were analyzed. Their effects on prognosis were analyzed by the Kaplan-Meier analysis, Log-rank test, Univariate and Multivariate Cox regression analysis. The potential biological function of RP11-757G1.5 in CRC was investigated by Colony formation, Edu cell proliferation, Flow cytometry, Wound healing and Transwell assays. Bioinformatics binding site analysis, Luciferase reporter assay, Ago2 immunoprecipitation assays, RNA pull-down assay, RT-qPCR and Western blotting were utilized to demonstrate the mechanism of RP11-757G1.5 acts as a molecular sponge of miR-139-5p to regulate the expression of YAP1. Finally, we further explore the potential role of RP11-757G1.5 in CRC orthotopic xenografts in vivio.Results: We discovered a novel oncogenic lncRNA RP11-757G1.5, that was overexpressed in CRC tissues, especially in aggressive cases. Moreover, up-regulation of RP11-757G1.5 strongly correlated with poor clinical outcomes of patients with CRC. Functional analyses revealed that RP11-757G1.5 promoted cell proliferation in vitro and in vivo. Furthermore, RP11-757G1.5 stimulated cell migration and invasion in vitro and in vivo. Mechanistic studies illustrated that RP11-757G1.5 regulated the expression of YAP1 through sponging miR-139-5p and inhibiting its activity thereby promoting CRC progression and development.Conclusions: Altogether, these results reveal a novel RP11-757G1.5/miR-139-5p/YAP1 regulatory axis that participates in CRC carcinogenesis and progression.

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