1.MiR-744-5p inhibits the proliferation, invasion, and migration of clear-cell renal cell carcinoma cells by targeting CCND1.
Kun Yang LEI ; Wen Jie XIE ; Ting SUN ; Yi Fu LIU ; Xu WANG
Journal of Southern Medical University 2022;42(5):712-717
OBJECTIVE:
To explore the role of miR-744-5p/CCND1 axis in clear-cell renal cell carcinoma (ccRCC).
METHODS:
We examined the expression levels of miR-744-5p in 65 pairs of ccRCC and adjacent tissue specimens and in 5 ccRCC cell lines and human renal tubular epithelial (HK2) cells using qRT-PCR. The ccRCC cell lines 786-O and OSRC2 were transfected with miR-744-5p mimic, CCND1 mimic, or their negative control mimics, and the changes in cell proliferation, migration, and invasion were evaluated with CCK-8, wound healing, and Transwell assays. The downstream target molecules of miR-744-5p were predicted by bioinformatics analysis, and the expression level of CCND1 in ccRCC cells was verified by qRT-PCR and Western blotting. The relationship between miR-744-5p and CCND1 was further validated by dual luciferase reporter assay, and the role of the miR-744-5p/CCND1 axis in ccRCC was explored by rescue experiments.
RESULTS:
MiR-744-5p was significantly downregulated in ccRCC tissues and cell lines (all P < 0.05), and its overexpression inhibited the proliferation, migration, and invasion of ccRCC cells (all P < 0.05). Bioinformatics analysis and dual luciferase reporter assay showed that CCND1 was a downstream target of miR-744-5p. The results of rescue experiments showed that upregulation of CCND1 could partially reverse the inhibitory effect of miR-744-5p overexpression on ccRCC cell proliferation, migration, and invasion (all P < 0.05).
CONCLUSION
MiR-744-5p inhibits the malignant phenotype of ccRCC cells by targeting CCND1, and the miR-744-5p/CCND1 axis may be a novel target for diagnosis and treatment of ccRCC.
Carcinoma, Renal Cell/metabolism*
;
Cell Line, Tumor
;
Cell Movement/genetics*
;
Cell Proliferation/genetics*
;
Cyclin D1/genetics*
;
Humans
;
Kidney Neoplasms/metabolism*
;
MicroRNAs/metabolism*
2.Expression of human telomerase reverse transcriptase in renal cell carcinoma and its clinical significance.
Peng WU ; Shao-bin ZHENG ; Yi ZUO ; Wan-long TAN ; Chen YAO ; Yao-dong JIANG
Journal of Southern Medical University 2008;28(2):272-274
OBJECTIVETo investigate the expression of human telomerase reverse transcriptase (hTERT) in renal cell carcinoma (RCC) and its clinical significance.
METHODSThe expression levels of hTERT mRNA and protein were detected using RT-RCR and Western blotting in 45 RCC tissues, 45 adjacent tissues and 786-0 cell line, and the associations of hTERT expression with the tumor size, clinical stage, pathological type and grade were evaluated.
RESULTShTERT mRNA and protein was expressed at significantly higher levels in RCC tissues than in the adjacent tissues (P=0.000), and no correlation of hTERT expression was found with the tumor size, clinical stage, pathological type or grade.
CONCLUSIONhTERT might serve as a diagnostic and prognostic marker for RCC, and also shed light on the new clues for gene therapy of RCC.
Adult ; Aged ; Biomarkers, Tumor ; genetics ; metabolism ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Telomerase ; genetics ; metabolism
3.Pathologic features of recently identified renal cell carcinoma.
Xiang FAN ; Qiu RAO ; Li-hua ZHANG
Chinese Journal of Pathology 2013;42(8):569-573
Adenocarcinoma, Follicular
;
genetics
;
metabolism
;
pathology
;
Carcinoma, Papillary
;
genetics
;
metabolism
;
pathology
;
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
pathology
;
Diagnosis, Differential
;
Humans
;
Immunohistochemistry
;
Kidney Diseases, Cystic
;
genetics
;
metabolism
;
pathology
;
Kidney Neoplasms
;
genetics
;
metabolism
;
pathology
;
Thyroid Neoplasms
;
genetics
;
metabolism
;
pathology
;
Translocation, Genetic
4.MiR-144 inhibits cell proliferation of renal cell carcinoma by targeting MTOR.
Cheng XIANG ; Shi-peng CUI ; You KE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2016;36(2):186-192
MicroRNAs (miRNAs) modulate the expression of tumorigenesis-related genes and play important roles in the development of various types of cancers. It has been reported that miR-144 is dysregulated and involved in multiple malignant tumors, but its role in renal cell carcinoma (RCC) remains elusive. In this study, we demonstrated miR-144 was significantly downregulated in human RCC. The decreased miR-144 correlated with tumor size and TNM stage. Moreover, overexpression of miR-144 in vitro suppressed RCC cell proliferation and G2 transition, which were reversed by inhibition of miR-144. Bioinformatic analysis predicted that mTOR was a potential target of miR-144, which was further confirmed by dual luciferase reporter assay. Additionally, the examination of clinical RCC specimens revealed that miR-144 was inversely related to mTOR. Furthermore, knocking down mTOR with siRNA had the same biological effects as those of miR-144 overexpression in RCC cells, including cell proliferation inhibition and S/G2 cell cycle arrest. In conclusion, our results indicate that miR-144 affects RCC progression by inhibiting mTOR expression, and targeting miR-144 may act as a novel strategy for RCC treatment.
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
pathology
;
Case-Control Studies
;
Cell Line, Tumor
;
Cell Proliferation
;
Female
;
G2 Phase
;
Humans
;
Kidney Neoplasms
;
genetics
;
metabolism
;
pathology
;
Male
;
MicroRNAs
;
genetics
;
metabolism
;
Middle Aged
;
S Phase
;
TOR Serine-Threonine Kinases
;
genetics
;
metabolism
5.Research advance in tumors associated with microphthalmia-associated transcription factor gene family.
Chinese Journal of Pathology 2011;40(7):496-498
Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
;
genetics
;
metabolism
;
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
Cell Cycle Proteins
;
genetics
;
metabolism
;
Humans
;
Melanoma
;
genetics
;
metabolism
;
Microphthalmia-Associated Transcription Factor
;
genetics
;
metabolism
;
Neoplasm Proteins
;
genetics
;
metabolism
;
Oncogene Proteins, Fusion
;
genetics
;
metabolism
;
Perivascular Epithelioid Cell Neoplasms
;
genetics
;
metabolism
;
Sarcoma, Clear Cell
;
genetics
;
metabolism
;
Translocation, Genetic
6.Read-through circular RNA rt-circ-HS promotes hypoxia inducible factor 1α expression and renal carcinoma cell proliferation, migration and invasiveness.
Yun Yi XU ; Zheng Zheng SU ; Lin Mao ZHENG ; Meng Ni ZHANG ; Jun Ya TAN ; Ya Lan YANG ; Meng Xin ZHANG ; Miao XU ; Ni CHEN ; Xue Qin CHEN ; Qiao ZHOU
Journal of Peking University(Health Sciences) 2023;55(2):217-227
OBJECTIVE:
To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.
METHODS:
Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.
RESULTS:
We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.
CONCLUSION
The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.
Humans
;
Carcinoma, Renal Cell/pathology*
;
Cell Proliferation
;
Hypoxia
;
Kidney Neoplasms
;
MicroRNAs/genetics*
;
Neoplasm Invasiveness/genetics*
;
RNA, Circular/metabolism*
;
RNA, Small Interfering
;
Hypoxia-Inducible Factor 1, alpha Subunit/genetics*
7.Expression of beta-human chorionic gonadotropin genes in renal cell cancer and benign renal disease tissues.
Yongguang JIANG ; Fuqing ZENG ; Chuanguo XIAO ; Junmin LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(3):291-293
To study the expression of beta-human chorionic gonadotropin (beta hCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of beta hCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for beta hCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of beta hCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. beta 7 was most frequently transcribed subtype gene independent on the histology. These findings suggested beta hCG gene transcription is not only involved in RCC but also in benign renal diseases.
Adult
;
Biomarkers, Tumor
;
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
Chorionic Gonadotropin, beta Subunit, Human
;
biosynthesis
;
genetics
;
Humans
;
Kidney
;
metabolism
;
Kidney Diseases
;
genetics
;
metabolism
;
Kidney Neoplasms
;
genetics
;
metabolism
;
Polycystic Kidney Diseases
;
metabolism
;
RNA, Messenger
;
biosynthesis
;
genetics
;
Reverse Transcriptase Polymerase Chain Reaction
8.Tumor suppressor gene VHL, hypoxia inducible factor, and renal cell carcinoma.
Yu-tao ZHANG ; Ni CHEN ; Hao ZENG ; Qiao ZHOU
Chinese Journal of Pathology 2006;35(9):562-564
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
pathology
;
Humans
;
Hypoxia-Inducible Factor 1, alpha Subunit
;
genetics
;
metabolism
;
Kidney Neoplasms
;
genetics
;
metabolism
;
pathology
;
Loss of Heterozygosity
;
Mutation
;
Von Hippel-Lindau Tumor Suppressor Protein
;
genetics
;
metabolism
;
von Hippel-Lindau Disease
;
genetics
;
metabolism
;
pathology
9.Long non-coding RNA MEG3 induces renal cell carcinoma cells apoptosis by activating the mitochondrial pathway.
Miao WANG ; Tao HUANG ; Gang LUO ; Chao HUANG ; Xing-Yuan XIAO ; Liang WANG ; Guo-Song JIANG ; Fu-Qing ZENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(4):541-545
This study aimed to examine the effect of long non-coding RNA (LncRNA) MEG3 on the biological behaviors of renal cell carcinoma (RCC) cells 786-0 and the possible mechanism. MEG3 expression levels were detected by RT-qPCR in tumor tissues and adjacent non-tumor tissues from 29 RCC patients and in RCC lines 786-0 and SN12 and human embryonic kidney cell line 293T. Plasmids GV144-MEG3 (MEG3 overexpression plasmid) and GV144 (control plasmid) were stably transfected into 786-0 cells by using lipofectamine 2000. Cell viabilities were determined by MTT, cell apoptosis rates by flow cytometry following PE Annexin V and 7AAD staining, apoptosis-related protein expressions by Western blotting, and Bcl-2 mRNA by RT-qPCR in the transfected cells. The results showed that MEG3 was evidently downregulated in RCC tissues (P<0.05) and RCC cell lines (P<0.05). The viabilities of 786-0 cells were decreased significantly after transfection with GV144-MEG3 for over 24 h (P<0.05). Consistently, the apoptosis rate was significantly increased in 786-0 cells transfected with GV144-MEG3 for 48 h (P<0.05). Furthermore, overexpression of MEG3 could reduce the expression of Bcl-2 and procaspase-9 proteins, enhance the expression of cleaved caspase-9 protein, and promote the release of cytochrome c protein to cytoplasm (P<0.05). Additionally, Bcl-2 mRNA level was declined by MEG3 overexpression (P<0.05). It was concluded that MEG3 induces the apoptosis of RCC cells possibly by activating the mitochondrial pathway.
Apoptosis
;
Carcinoma, Renal Cell
;
genetics
;
metabolism
;
pathology
;
Cell Line, Tumor
;
Cell Survival
;
Gene Expression Regulation, Neoplastic
;
HEK293 Cells
;
Humans
;
Kidney Neoplasms
;
genetics
;
metabolism
;
pathology
;
Mitochondria
;
genetics
;
RNA, Long Noncoding
;
genetics
;
metabolism
;
Signal Transduction
10.Reversal of multidrug resistance in renal cell carcinoma by short hairpin RNA targeting MDR1 gene.
Yi-Xin HAO ; Zheng-Wen HE ; Jian-Hua ZHU ; Qian SHEN ; Jun-Zhong SUN ; Nan DU ; Wen-Hua XIAO
Chinese Medical Journal 2012;125(15):2741-2745
BACKGROUNDOver-expression of P-glycoprotein (P-gp), encoded by the MDR1 gene, confers multidrug resistance (MDR) in renal cell carcinoma (RCC) and is a major reason for unsuccessful chemotherapy. This study aimed to determine the effct of RNA interference (RNAi) on the reversal of MDR in human RCC.
METHODSWe designed and selected one short hairpin RNA (shRNA) targeting MDR1 gene, which is stably expressed from integrated plasmid and transfected by lentivirus fluid in human RCC A498 cell.
RESULTSThe MDR1-targeted RNAi resulted in decreased MDR1 gene mRNA level (P < 0.001), almost abolished P-gp expression and reversed MDR to different chemotherapy drugs in the RCC A498 cell line.
CONCLUSIONMDR could be reversed by RNAi in human RCC A498 cell line, which may be used for clinical application in future.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Carcinoma, Renal Cell ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Inhibitory Concentration 50 ; Lentivirus ; genetics ; RNA, Small Interfering ; genetics ; metabolism