ABSTRACT
OBJECTIVE: To study the effect of arsenic trioxide (As2O3) and all-trans retinoic acid (ATRA) on human cervical carcinoma HeLa cell line. METHODS: HeLa cells were treated with As2O3 and ATRA. The cell proliferation was evaluated by MTT assay. The expressions of NDRG-1 protein and mRNA were determined by Western blot and RT-PCR analysis. RESULTS: MTT assay showed that As2O3 and ATRA inhibited the growth of human cervical carcinoma HeLa cells in vitro in a dose- and time-dependent manner. Western blot and RT-PCR techniques showed that As2O3 and ATRA down-regulated the expressions of NDRG-1 protein and mRNA (P < 0.05). CONCLUSION: As2O3 and ATRA can significantly inhibit the growth and proliferation of HeLa cells. The reason of these changes may be related with the down-regulation of expression of NDRG-1.
Subject(s)
Antineoplastic Agents/pharmacology , Arsenicals/pharmacology , Cell Cycle Proteins/metabolism , Cell Proliferation/drug effects , Intracellular Signaling Peptides and Proteins/metabolism , Oxides/pharmacology , Tretinoin/pharmacology , Antineoplastic Agents/administration & dosage , Arsenic Trioxide , Arsenicals/administration & dosage , Blotting, Western , Cell Cycle Proteins/genetics , Dose-Response Relationship, Drug , HeLa Cells , Humans , Intracellular Signaling Peptides and Proteins/genetics , Oxides/administration & dosage , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tretinoin/administration & dosageABSTRACT
BACKGROUND & OBJECTIVE: Survivin gene overexpresses in a variety of human tumors, and plays an important role in cell apoptosis and drug resistance of tumors. This study was designed to establish Survivin antisense RNA, and explore its effects on apoptosis of ovarian cancer cell line SKOV3 and sensitivity to docetaxel. METHODS: Survivin antisense eukaryotic expression vector anti-pcDNA3-svv was established, and transfected into SKOV3 cells by electroperforation. Positive clones (SKOV3-SVVanti) were screened out. Survivin mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR); Survivin protein was detected by Western blot. The effect of Survivin antisense RNA on apoptosis of SKOV3 cells was measured by flow cytometry and observed under electron microscope; its effect on sensitivity of SKOV3 cells to docetaxel was examined by MTT assay. RESULTS: The mRNA and protein levels of Survivin were obviously lower in SKOV3-SVVanti cells than in control cells. Terminal apoptosis changes were observed under electron microscope after transfection. The apoptosis rate was 19%. The 50% inhibitory concentration (IC50) of docetaxel was significantly lower for SKOV3-SVVanti cells than for control cells [(13.3+/-2.2) ng/ml vs. (53.2+/-2.4) ng/ml, P<0.05]. CONCLUSION: Surivivin antisense RNA can induce apoptosis of SKOV3 cells, and sensitize SKOV3 cells to docetaxel.
Subject(s)
Apoptosis , Microtubule-Associated Proteins/biosynthesis , Ovarian Neoplasms/pathology , RNA, Antisense/genetics , Taxoids/pharmacology , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Docetaxel , Drug Resistance, Neoplasm/drug effects , Female , Gene Expression Regulation, Neoplastic , Humans , Inhibitor of Apoptosis Proteins , Inhibitory Concentration 50 , Microtubule-Associated Proteins/genetics , RNA, Messenger/metabolism , Survivin , TransfectionABSTRACT
Structural role of the second copy of the rod-core linker CpcG, which was found by genome analysis, was studied in Synechocystis sp. PCC 6803 by gene disruption and fractionation of phycobilisome (sub)complexes. Disruption of cpcG2 (sll1471) resulted in a marked decrease in phycocyanin content both in the background of wild-type and cpcG1 (slr2051)-disruptant. The unique phycocyanin rod-CpcG2 complex without the major allophycocyanin components was isolated from the cpcG1-disruptant. By fluorescence analysis, it was proposed that CpcG2 protein connects the rods with a minor allophycocyanin component, to support energy transfer to Photosystem I.
