摘要
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 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.
