Experimental Study Of¹³¹I-K237Preparation And Its Effect Of Prostate Cancer Cells In Vitro

Objective: To explore the¹³¹I labeled peptides K237，get radiochemical purity higherthan95%of the¹³¹I-K237best conditions，Study The labeled product stability and biologicalactivity in vitro. Identification of¹³¹I-K237role on the treatment of prostate cancer LNCaPcells. in vitro.Methods:¹³¹I labeled K237with Iodogen method, By orthogonal experimental screeningbest mark condition. Using high performance liquid chromatograph （HPLC） combined withthin layer chromatography （TLC） Determination of labeling rate and radiochemical purity.Using Sephadex G25column chromatography separation and purification of¹³¹I-K237.¹³¹I-K237（radiochemical purity>95%） placed at room temperature and37℃water bathrespectively, measure its radiochemical purityfor Observing its stability in a different time. Tocomplete Human umbilical vein endothelial cells （HUVEC） proliferation inhibitionexperiments with CCK-8methed. To determine the¹³¹I-K237biological activitiesBycalculating K237and¹³¹I-K237inhibition rate of HUVEC. Prostate cancer LNCaP cellscultured in vitro, the different radioactivity of¹³¹I-K237, the same concentration K237andblank control group intervention48h, the application of optical microscopy, fluorescencemicroscopy, nucleic acid gel electrophoresis and flow cytometry analysis to identify the roleof¹³¹I-K237treatment of LNCaP cells.Results: The best labeling conditions: K237100μg,Iodogen50μg and by the reactiontime of40min at37℃,the label rate reached up to70%, By the Sephadex G25columnchromatography separation and purification, the radiochemical purity can reach more than95%. Stability experiment results show that¹³¹I-K237was placed24h, its radiochemical purity>90%.There was no statistically significant difference on the¹³¹I-K237and K237onHUVEC cell proliferation inhibition rate difference （P>0.05）, K237biological activities didnot change after¹³¹I labeling. Role of¹³¹I-K237for the treatment of LNCaP cells: Lightmicroscope can be observed morphology of apoptotic cells,apoptotic cells was showedclumps or fragmental strong blue fluorescence with fluorescent light microscope, DNAelectrophoresis is a typical "ladder" stripe, Flow cytometry analysis results:¹³¹I-K237andK237effect on LNCaP cells capable of inducing apoptosis, But the¹³¹I-K237’s effect isobvious higher than K237’s.Conclusion:¹³¹I-labeled peptide K237by the Iodogen method is simple and efficient,the biological activity of the labeled product was preserved, and was not obviously damaged..¹³¹I-K237treatment LNCaP cells cells have a significant effect, It is a foreshadowing foranimal experiment and clinical application in the late.