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This work was undertaken to gain further information on the molecular mechanismsunderlying autophagosome formation and its relation with tumor cell survival inresponse to radiation in colon cancer. A human colon cancer cell line, HCT-116,was examined with respect to cell survival after blockade of irradiation-inducedautophagosome formation by pharmacological interference. Autophagosome formationwas confirmed using a kinetic study with incorporated bovine serum albumin gold-conjugate(BSA-Au) analyzed by electron microscopy and an autophagosome-associated LC3Bantibody measured by immunofluorescence and Western blotting. Annexin V/PI doublestaining was used to monitor cell death by apoptosis, and cell cycle profilesby flow cytometry. Ionizing radiation (IR) promoted autophagosome formation inthe HCT-116 IR-surviving cells. Pharmacological interference showed that PI3K/Aktand Src were involved in early stages of autophagosome formation. IR alone decreasedcell proliferation by arresting cells in the G2/M phase, and pharmacological interferenceof autophagosome formation decreased proliferation, but did not affect cell survival.Also, our data suggest that decreased proliferation caused by PI3K and Src inhibitorscould be through S phase cell cycle delay. Our results clearly indicate that blockadeof IR-induced autophagosome formation impairs proliferation but does not enhancecell death in colon cancer cells.

international journal of oncology

Blockade of irradiation-induced autophagosome formation impairs proliferation but does not enhance cell death in HCT-116 human colorectal carcinoma cells