Repression Of EGFR And Induction Of Apoptosis In Human Ovarian Cancer SKOV3/DDP Cells By SiRNA Silenced HDAC1Gene

Ovarian cancer is one of the malignant tumors which seriously threaten towomen’s health,and the mortality is so high.So far, drug resistance of cancer is stillthe main cause of the chemotherapy failure. Looking for a better and safer way toovercome the drug resistance of cancer is a difficuty problem all the way. This article,looking for the highly expressed genes of ovarian cance named EGFR gene which hasbeen a new target for cancer treatment.RNA interference (RNAi) technology exploresthe influence of EGFR expression and the cell cycle and apoptosis in ovarian cancer.It will provide a reliable clinical basis for reversal of tumor drug resistance.It is reported that ultrasound microbubble auxiliary RNAi technology maybecome the important means to attack the ovarian cancer. Ultrasound microbubblemainly increases the rate of gene transfection. So in vitro, only the transfection ratereaches a certain value, can it illustrate interference effects. RNA interference (RNAi)is a means of gene silencing,it is different from knockout,it is based on no DNAdamage,interfere with some gene, make the gene expression reduce or disappear andthe status stably expression in cells.The gene silence is related to tumor occurrenceand tumor multi-resistant, it has becoming the new trend for the treatment of cancer,there has a report: gene silencing related to acetylation. the core group proteinacetylation level of each area of chromosomes is restricted by the histone deacetylaseand histone acetyltransferase which play the dynamic balance roles, when the histoneacetyl-transferase has become the main action, transcription activity is activated, onthe contrary, transcription activity will be inhibited. When this balance is broken, thecells will worsen, it will cause cancer.Studies have shown that EGFRinhibitors,especially its drug-resistant problem of epithelial tumors’ clinical treatment,many clinical studies have proved that it’s associated with EGFR expressionand high drug-resistance, Studies have shown that HDACs is crucial in EGFRtranscription expression, HDACi did not affect the degradation of EGFR mRNA incolorectal cancer cells. However in ER-negative human breast cancer, HDACidecreased the EGFR mRNA stability, HDACi disrupt the signaling pathway viasilencing EGFR expression,so we guess the fact that HDACi can’t be used in all theteatment of cancers. According to this fact the relationship of HDAC1and EGFR ischecked in ovarian cancer of drug-resistance, combining ligands of EGFR with EGFRfamily form the different homogeneous or heterogeneous dimer which can buildcomplicated biochemical pathways, the main path ways of EGFR are theRas/Raf/MEK/ERK and PI3K/Akt/PDK1. SP1is essential to EGFR transcription,HDACIs can separate SP1from EGFR to reduce the EGFR expression. BothCisplatin and taxol are recognized as the most effective treatment of ovarian cancerchemotherapy drugs.In this article,we choose cisplatin-sensitive SKOV3and theircisplatin-resistant clones SKOV3/DDP as the in vitro model, targeting importsiRNA-HDAC1to show EGFR gene expression regulation in SKOV3/DDP resistantcells.Expected results for siRNA-HDAC1can improve sensitivity of ovarian cancerchemotherapy,it is essential in reversing ovarian cancer cell lines cisplatin resistance.The experiments as follows:The methods of part Ⅰmainly illustrate the difference research of HDAC1expression in ovarian cancer cells:the cell viability was detected by MTT assay andIC50.The level of HDAC1protein was detected by using immunocytochemicalstaining in ovarian cancer cells.Results:SKOV3/DDP cells resistance index is3.2.Thelevel of HDAC1expression in SKOV3/DDP cells is higher than in SKOV3cells.The methods of part Ⅱmainly illustrate the influence of interference plasmid onthe growth of ovarian cancer cells:the plasmid was sent to the company sequencingand proved successful extraction of plasmid.The expression level of EGFR mRNAand HDAC1mRNA and protein was analyzed by Real-time RCR and cell viabilitywas detected by MTT assay.Cell cycle and apoptisis was detected by flow cytometry. Results: Recombinant plasmid was successfully extracted,and it can be used fortransfection.The level of EGFR mRNA reduced after transfection of HDAC1reconversion plasmid;Flow cytometry shows that the cycle of transfected cells wasarrested in G0/G1phase. The apoptisis will be induced after transfection of HDAC1reconversion plasmid.Conclusions:1.HDAC1and EGFR are overexpressed in ovarian cancer cells of drugresistance.The expression is associated with tumor MDR.2.siRNA-HDAC1was transfected into ovarian cancer cells can decrease theEGFR expression lever of mRNA.Surmising EGFR is involved in cell regulation ofcell cycle process and apoptosis.At the same time siRNA-HDAC1interferencepromotes cancer cell apoptosis.3.siRNA-HDAC1can increase the sensitivity of the ovarian cancer cells todrugs,the tageted therapy of HDAC1may apply to the treatment of the drug-resistantovarian cancer,reversal the drug-resistant.