Rotavirus induces apoptosis in fully differentiated human intestinal Caco-2 cells.
Rotaviruses, which are the main cause of viral gastroenteritis in young children, induce structural and functional damages in infected mature enterocytes of the small intestine. To investigate a relationship between rotavirus infection and cell death by apoptosis, we used the human intestinal Caco-2 cell line. We demonstrated by several methods including TUNEL and ELISA detection of cytoplasmic histone-associated DNA fragments that the infection of fully differentiated Caco-2 cells by the RRV rotavirus strain induces apoptosis. Rotavirus infection leads to the loss of mitochondrial membrane potential and the release of cytochrome C from mitochondria. We showed that rotavirus-induced apoptosis was dependent of the multiplicity of infection and increased with time from 4 h to 24 h of infection. Flow cytometric analysis showed that DNA fragmentation occurs in productively infected cells, suggesting that rotavirus induces apoptosis by a direct mechanism. We also demonstrated that non-replicative RRV particles are not sufficient to induce apoptosis and viral gene expression seems required. Intracellular calcium plays a role in RRV-induced apoptosis because treatment with an intracellular calcium ion chelator (BAPTA-AM) partially inhibited apoptosis.
Chaïbi
Charlotte
C
Institut National de la Santé et de la Recherche Médicale, U 510, Pathogènes et Fonctions des Cellules Epithéliales Polarisées, Faculté de Pharmacie, 5 rue J.B. Clément, 92290 Châtenay-Malabry, France.
Cotte-Laffitte
Jacqueline
J
Sandré
Catherine
C
Esclatine
Audrey
A
Servin
Alain L
AL
Quéro
Anne-Marie
AM
Géniteau-Legendre
Monique
M
D000230:Adenocarcinoma; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms; D045304:Cytochromes c / Q000032:analysis; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D008564:Membrane Potentials / Q000502:physiology; D008928:Mitochondria / Q000502:physiology; D012401:Rotavirus / Q000472:pathogenicity*
D017304:Annexin A5; D045304:Cytochromes c