Hypoxia and Serum deprivation protected MiaPaCa-2 cells from KAI1-induced proliferation inhibition through autophagy pathway activation in solid tumors

Purpose. KAI1 closely correlates with pancreatic cancer metastasis. There might be some factors that protect the cells from a proliferation inhibition by KAI1 in the solid tumors’ microenvironment. Hypoxia and ischemia are the main characteristics of the microenvironment within solid tumors. Whether they affect the KAI1 inhibitory effects on cell proliferation is still unclear. Methods. MiaPaCa-2 human pancreatic cancer cells do not express KAI1 protein. However, after being infected with Ad5-KAI1, they expressed KAI1 protein. We cultured them under hypoxic and serum-free conditions to simulate the solid tumor hypoxic-ischemic microenvironment. The cells were divided into the control, hypoxic, serum-free, and hypoxic with serum-free groups. The proliferation and apoptosis were observed by CCK8 and Annexin V-FITC/PI, respectively. The green fluorescent protein-labeled light chain 3 association with autophagosome membranes was detected by confocal microscopy. The ratio of LC3-II–LC3-I expression level was detected by western blot. Pretreatment of 3-MA was used to inhibit the autophagy. We, then observed whether the hypoxic and serum-free conditions could change the effect of KAI1 on cell survival and whether the pretreatment of 3-MA could inhibit the effect of hypoxic and serum-free conditions on KAI1 function. Results. Hypoxia and serum-free media effectively reduced the apoptosis and proliferation inhibition caused by KAI1 and was beneficial to the cell survival. 3-MA pretreatment effectively blocked the protective effect of hypoxia and serum-free media on the cells by autophagy block. Conclusions. Serum-free media and hypoxia protected the MiaPaCa-2 cells from a KAI1-induced apoptosis and proliferation inhibition via autophagy induction (AU)

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Deteccion y características de células prostáticas circulantes primarias, asociación con la presencia de micrometástasis y las implicaciones para el tratamiento quirúrgico en hombres con cáncer prostático. El estudio ProTECT (Prostate Tumor Early Cancer Test) / Detection and characteristics of primary circulating prostate cells; association with micrometastasis and implications for surgical treatment of men with prostate cáncer

OBJETIVO: Determinar la frecuencia de células prostáticas circulantes (CPCs) primaria en hombres con cáncer de próstata en el momento del diagnóstico, la asociación con micrometástasis ósea y subclasificación por CD82. Determinar la relación con la estadio patológica y la eficacia para seleccionar pacientes para la prostatectomía radical.MÉTODOS: Se incluyeron hombres con diagnóstico de cáncer de próstata previo a tratamiento definitivo. Se obtuvieron muestras de sangre y de médula ósea, las células mononucleares separadas por centrifugación diferencial y células prostáticas identificadas con inmumocitoquímica con anti-APE, las muestras positivas fueron sub-clasificadas con anti-CD82. Se registraron también los detalles de APE sérico, Índice de Gleason y estadio patológica.RESULTADOS: De 77 hombres, 58 (75,3%) tuvieron 1° CPCs detectadas. Hubo una asociación con estadio pero no con el Índice de Gleason, 31 (40,3%) tuvieron micrometástasis. Hubo una asociación significativa con la estadio patológica y Índice de Gleason. Pacientes CPC negativa tuvieron una menor frecuencia de micrometástasis que los hombres CPC positiva 1/19 versus 30/58 (p<0,0003).Hubo una relación inversa significativa entre la expresión de CD82 en CPCs y el índice de Gleason y menor frecuencia de micrometástasis en comparación con hombres CPC CD82 positivos (p<0,0005).En el grupo de combinación de hombres CPC negativa y CPC positiva CD82 positivo la frecuencia de micrometastasis fue significativamente menor que el grupo CPC (+) CD82 (-) 5/39 versus 26/38 respectivamente(p<0,0000007), con una sensibilidad de 87% y especificidad de 73,9% para la ausencia de micrometástasis(AU)

CONCLUSIONES: La presencia de CPCs implica un riesgo mayor de desarrollar micrometástasis, la co-expresión del CD82 es asociada con tumores de bajo grado, un riesgo disminuido del desarrollo de micrometástasis óseas. Como consecuencia, el uso de la detección de CPCs primarias y su sub-clasificación podrían ser clínicamente útiles para identificar los pacientes los cuales beneficiarán de una prostatectomía radical como tratamiento de primera línea(AU)

OBJECTIVES: To determine the frequency of primary circulating prostate cells in men with prostate cancer at the time of diagnosis, the association with micrometastasis, sub-classification for CD82 and the relation with pathological stage. To determine their clinical usefulness to identify patients in whom radical prostatectomy would be first choice therapy.METHODS: Men with the diagnosis of prostate cancer before definitive therapy. Blood and bone marrow samples were taken, mononuclear cells separated by differential centrifugation and prostate cells identified with immunocytochemistry using anti-PSA. Positive samples were sub-classified with anti-CD82. Details of serum PSA, Gleason score and pathological stage were registered.RESULTS: Of 77 men 58 (75.3%) had primary CPCs detected, there was an association with stage but not Gleason. 31 (40.3%) had micrometastasis with an association with stage and Gleason score. CPC-negative patients had fewer micrometastasis detected, 1/19 versus 30/58 (p<0.003).There was an inverse relation between CD82 expression and Gleason score, men with CPCs expressing CD82 had fewer micrometastasis. The combined group of CPC negative and CPC positive CD82 positive men showed a sensitivity of 87% and specificity of 73.9% for the absence of micrometastasis.CONCLUSIONS: The detection of CPCs and sub-classification with CD82 could be clinically useful to identify men with a significantly lower risk of micrometastais and as a consequence to identify men in whom radical prostatectomy could be the best initial treatment(AU)