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AIM:To investigate the expression and function of classicalprotein kinase C(PKC)isoenzymes in inducing MDRphenotype in gastric cancer cells.METHODS:Two cell lines were used in the study:gastriccancer cell SGC7901 and its drug-resistant cell SGC7901/VCRstepwise-selected by vincristine 0.3,0.7 and 1.0 mg·L~(-1),respectively.The expression of classical PKC(cPKC)isoenzymes in SGC7901 cells and SGC7901/VCR cells weredetected using immunofluorescent cytochemistry,laserconfocal scanning microscope and Western blot.The effectsof anti-PKC isoenzymes antibody on adriamycinaccumulation in SGC7901/VCR cells were determined usingflow cytometric analysis.RESULTS:(1)SGC7901 cells exhibited positive staining ofPKC-α.SGC7901/VCR cells exhibited stronger staining ofPKC-α than SGC790t cells.The higher dosage vincristineselected,the much stronger staining of PKC-α was observedon SGC7901/VCR cells.(2)Both SGC7901 and SGC7901/VCRcells exhibited positive staining of PKC-βⅠ and PKC-βⅡ withno significant difference.(3)Compared with SGC7901,SGC7901/VCR cells had decreased adriamycin accumulationand retention.Accumulation of adriamycin in SGC7901 was5.21±2.56mg·L~(-1),in SGC7901/VCR 0.3 was 0.85±0.29mg·L~(-1),in SGC7901/VCR 0.7 was 0.81±0.32mg·L~(-1),and inSGC7901/VCR 1.0 was 0.80±0.33mg·L~(-1);Retention ofadriamycin in SGC 7901 was 2.51±1.23mg·L~(-1),in SGC7901/VCR 0.3 was 0.47±0.14 mg·L~(-1),in SGC7901/VCR 0.7 was 0.44±0.15 mg·L~(-1),and in SGC 7901/VCR 1.0 was 0.41±0.11mg·L~(-1).(4)Fluorescence intensity presented adriamycinaccumulation in SGC7901/VCR cells was increased from 1.14±0.36 to 2.71±0.94 when cells were co-incubated with anti-PKC-αbut not with anti-PKC,-βⅠ,PKC-βⅡ and PKCγ antibodies. CONCLUSION:PKC-α,but not PKC-αⅠ,PKC-βⅡ or PKCγ,may play a role in multidrug resistance of gastric cancercells SGC7901/VCR.
AIM: To investigate the expression and function of classical protein kinase C (PKC) isoenzymes in inducing MDRphenotype in gastric cancer cells.METHODS: Two cell lines were used in the study:gastriccancer cell SGC7901 and its drug-resistant cell SGC7901/VCRstepwise-selected by Vincristine 0.3,0.7 and 1.0 mg·L-1,respectively.The expression of classical PKC(cPKC)isoenzymes in SGC7901 cells and SGC7901/VCR cells were detected using immunofluorescent cytochemistry,laser confocal scanning microscope and Western blot.The effects of anti- PKC isoenzymes antibody on adriamycinaccumulation in SGC7901/VCR cells were determined using flow cytometric analysis.RESULTS:(1)SGC7901 cells exhibited positive staining ofPKC-α.SGC7901/VCR cells exhibited strong staining ofPKC-α than SGC790t cells.The higher dosage vincristine selected,the Much powerful staining of PKC-α was observedon SGC7901/VCR cells. (2)Both SGC7901 and SGC7901/VCRcells exhibited positive staining of PKC-βI and PKC-βII withno signifi Cant difference. (3)Compared with SGC7901,SGC7901/VCR cells had decreased adriamycin accumulation and retention. Accumulation of adriamycin in SGC7901 was 5.21±2.56 mg·L -1, in SGC7901/VCR 0.3 was 0.85±0.29 mg· L~(-1), in SGC7901/VCR 0.7 was 0.81±0.32mg·L~(-1), and inSGC7901/VCR 1.0 was 0.80±0.33mg·L~(-1); Retention of adriamycin in SGC 7901 was 2.51 ±1.23 mg·L -1 in SGC7901/VCR 0.3 was 0.47±0.14 mg·L -1 in SGC7901/VCR 0.7 was 0.44±0.15 mg·L -1 and in SGC 7901/VCR 1.0 was 0.41±0.11 mg·L~(-1).(4) Fluorescence intensity gave adriamycinaccumulation in SGC7901/VCR cells was increased from 1.14±0.36 to 2.71±0.94 when cells were co-incubated with anti-PKC- Conclusions: PKC-α, but not PKC-αI, PKC-βII or PKCγ, may play a role in multidrug resistance of gastric cancer cells SGC7901/VCR.