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Objective:To investigatethesignaltransductionmechanismof gap junctionalgenesconnexin43in human cervicalcarcinogenesis.Methods:HumancervicalcarcinomacelllineHeLawas culturedandtreatedby all-trans-retinoic acid(ATRA).Flowcytometer(FCM)wasemployedto detectexpressionof Cx43proteininHeLacells.Fluo-3AM loadingandlaserscanningconfocalmicroscope(LSCM)wereusedto measuretheconcentrationsof intracellularcalcium([Ca 2+ ] i )in HeLacells.Phosphorylationon tyrosineof connexin43proteinwas examinedby immunoblot.Results:The positiverateof Cx43proteinincreasedfrom1.9%inuntreatedHeLacellsto26.3%inRA-treatedHeLacellsas shownby FCM.[Ca 2+ ] i was35.73nmol/Lin untreatedHeLacellswhichwas increasedto58.16nmol/Lin ATRA-treatedcells.ImmunoblotshowedthatATRA-treatedHeLacellshad phosphorylationon tyrosinein Cx43proteinwhereasuntreated cellshadnot.Conclusions:Carcinogenesisof humancervicalcarcinomais relatedwiththeabnormalexpressionof cx43geneand disorderof signaltransductionmanifestedas the decreaseof[Ca 2+ ] i and post-translationphosphorylationon tyrosineof Cx43protein.Theanti-tumoreffectof ATRAinHeLacellsmightbe dueto theup-regulationof cx43geneand itssignaltransductionpathway.
Objective: To investigatethesignal transductionmechanismof gap junctionalgenesconnexin43in human cervicalcarcinogenesis.Methods:HumancervicalcarcinomacelllineHeLawas culturedandtreatedbyall-trans-retinoic acid(ATRA).Flowcytometer(FCM)wasemployedto detectexpressionof Cx43proteininHeLacells.Fluo-3AM loadingandlaserscanningconfocalmicroscope(LSCM)wereusedto measuretheconcentrationsof intracellularcalcium([Ca 2+ ] i )in HeLacells.Phosphorylationon tyrosineof connexin43proteinwas examined by immunoblot.Results:The positiverateof Cx43proteinincreasedfrom 1.9%inuntreatedHeLacellsto26.3%inRA-treatedHeLacellsas shownby FCM.[Ca 2+ ]iwas35.73nmol/Lin untreatedHeLacellswhichwas increasedto58.16nmol/Lin ATRA-treatedcells.ImmunoblotshowedthatATRA-treatedHeLacellshad Phosphorylationon tyrosinein Cx43proteinwhereasuntreated cellshadnot.Conclusions:Carcinogenesisof humancervicalcarcinomais relatedwiththeabnormalexpressionof cx43geneand disorderof signaltransductionmanifestedas t He decreaseof [Ca 2+ ] i and post-translation phosphorylationon tyrosineof Cx43protein. Theanti-tumoreffectof ATRAinHeLacellsmightbe dueto theup-regulationof cx43geneand itssignaltransductionpathway.