1.Effects of anti-HPV16E6-ribozyme on phenotype and gene expression of a cervical cancer cell line.
Yanfang ZHENG ; Jiren ZHANG ; Lianghu QU
Chinese Medical Journal 2002;115(10):1501-1506
OBJECTIVETo investigate the effects of anti-HPV16E6-ribozyme (HRz) on phenotype and gene expression of a cervical cancer cell line.
METHODSHRz was designed by computer programs. HRz's activity was identified by cleavage experiments in vitro. HRz and empty eukaryotic plasmids were transfected into CaSKi cells with lipofectin, then renamed CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blot. The amounts of E6 mRNA in three kinds of cells lines were detected by Northern blot. Cell growth curves and soft agar forming ability were studied. The ability of each cell line to form tumors was assessed in nude mice. Apoptosis rates and expression of c-myc, bcl-2, p53 and Fas were detected by flow cytometry (FCM). Antigens of tumor cells, HLA-1, HLA-2, B7-1 and B7-2 were also detected. NK, LAK, and CD(3)AK cells were induced. Their cytotoxicities were detected in CaSKi-R, CaSKi-P, and CaSKi cells.
RESULTSIn vitro cleavage reaction demonstrated that HRz could cleave HPV16E6 mRNA in a site-specific manner. HRz could be expressed stably in transfected CaSKi cells. Northern blot analysis showed that E6 mRNA levels were lower in CaSKi-R than in CaSKi. The growth rate of CaSKi-R was slower than those of CaSKi and CaSKi-P. The soft agar-forming rate of CaSKi-R was lower compared with those of CaSKi and CaSKi-P cells. The ability of CaSKi-R to form tumors in nude mice was also poor. The apoptosis rate of CaSKi-R cells was much higher than those of CaSKi and CaSKi-P. HRz could reduce the expression of E6, c-myc and bcl-2 proteins, and increase the expression of p53 as well. HRz could increase the expression of HLA-2, B7-1 and B7-2 antigens. The cytotoxicity of NK, LAK and CD(3)AK cells was much higher in CaSKi-R than in CaSKi-P and CaSKi cells.
CONCLUSIONHRz not only reverses the malignant phenotype of CaSKi cells partially, but also induces apoptosis in the cells, and increases sensitivity of CaSKi cells to immune cells.
Animals ; Apoptosis ; Cell Division ; Cytotoxicity, Immunologic ; Female ; Flow Cytometry ; Gene Expression ; Genetic Therapy ; Humans ; Killer Cells, Natural ; immunology ; Mice ; Mice, Nude ; Oncogene Proteins, Viral ; genetics ; Phenotype ; RNA, Catalytic ; therapeutic use ; RNA, Messenger ; analysis ; Repressor Proteins ; Tumor Cells, Cultured ; Uterine Cervical Neoplasms ; genetics ; pathology ; therapy
2.Applications of RNA Indexes for Precision Oncology in Breast Cancer.
Liming MA ; Zirui LIANG ; Hui ZHOU ; Lianghu QU
Genomics, Proteomics & Bioinformatics 2018;16(2):108-119
Precision oncology aims to offer the most appropriate treatments to cancer patients mainly based on their individual genetic information. Genomics has provided numerous valuable data on driver mutations and risk loci; however, it remains a formidable challenge to transform these data into therapeutic agents. Transcriptomics describes the multifarious expression patterns of both mRNAs and non-coding RNAs (ncRNAs), which facilitates the deciphering of genomic codes. In this review, we take breast cancer as an example to demonstrate the applications of these rich RNA resources in precision medicine exploration. These include the use of mRNA profiles in triple-negative breast cancer (TNBC) subtyping to inform corresponding candidate targeted therapies; current advancements and achievements of high-throughput RNA interference (RNAi) screening technologies in breast cancer; and microRNAs as functional signatures for defining cell identities and regulating the biological activities of breast cancer cells. We summarize the benefits of transcriptomic analyses in breast cancer management and propose that unscrambling the core signaling networks of cancer may be an important task of multiple-omic data integration for precision oncology.
Female
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Gene Expression Profiling
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Genomics
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Humans
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MicroRNAs
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metabolism
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Precision Medicine
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RNA Interference
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RNA, Messenger
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metabolism
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Triple Negative Breast Neoplasms
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classification
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genetics
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metabolism
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therapy