Histochemical, immunohistochemical and ultrastructural studies were performed on cases of hepatocellular carcinoma (HCC) with pale bodies (PB). HCC containing PBs was observed in 3 (5.5%) of 55 consecutively resected HCC cases. Histologically, a large number of hepatocytes displayed pale or eosinophilic staining of the cytoplasm, resulting in ground-glass appearance. They were aggregated in nodular pattern, or diffusely intermixed with other malignant hepatocytes. PBs were negative for periodic-acid Schiff and Masson's trichrome staining. The inclusions showed a strong positive reaction for fibrinogen and some of them were weakly positive for albumin but negative for hepatitis B surface antigen, hepatitis B core antigen, alpha-fetoprotein and alpha-1-antitrypsin. Ultrastructurally, PBs were membrane-bound and contained granular materials of moderate electron density, and were closely related to dilated rough endoplasmic reticulum. These findings support that PBs are secretory fibrinogen accumulated in cystic ER and that such intracellular accumulation possibly reflects a defective transport of fibrinogen.
Albumins/analysis ; Carcinoma, Hepatocellular/pathology* ; Cytoplasm/ultrastructure ; Cytoplasm/pathology ; Cytoplasm/chemistry ; Endoplasmic Reticulum, Rough/ultrastructure ; Endoplasmic Reticulum, Rough/pathology ; Endoplasmic Reticulum, Rough/chemistry ; Fibrinogen/analysis ; Human ; Inclusion Bodies/ultrastructure ; Inclusion Bodies/pathology* ; Inclusion Bodies/chemistry ; Liver Neoplasms/pathology* ; Male ; Microscopy, Electron ; Middle Age ; Periodic Acid-Schiff Reaction

OBJECTIVETo observe the ultrastructural changes of HeLa cells in response to tubeimoside I (TBMS1) treatment and the protective effect of cyclosporine A (CsA), and explore the role of intrinsic apoptosis pathway in TBMS1-induced HeLa cell apoptosis.

METHODSHeLa cells were treated with TBMS1 (10-50 micromo/L) alone or in combination with 2 micromol/L CsA for 12 and 24 h and observed with transmission electron microscope (TEM) for the ultrastructural changes of the cells.

RESULTSTBMS1 induced apoptosis of HeLa cells in a concentration- and time-dependant manner. Under TEM, the treated cells progressively shrunk and the intercellular space widened with loss of microvillus, mitochondrial swelling, rough endoplasmic reticulum enlargement, chromatin condensation, nuclear shrinkage and nuclear pyknosis as TBMS1 concentration increased. At low concentrations, CsA offered partial protection of the mitochondria from TBMS1-induced damage whereas high-concentration CsA did not.

CONCLUSIONTBMS1 induces ultrastructural changes typical for apoptosis of the HeLa cells, which provides morphological evidence for the role of intrinsic apoptosis pathway in TBMS1-induced apoptosis.

Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cyclosporine ; pharmacology ; Dose-Response Relationship, Drug ; Endoplasmic Reticulum, Rough ; drug effects ; ultrastructure ; Female ; HeLa Cells ; Humans ; Immunosuppressive Agents ; pharmacology ; Microscopy, Electron, Transmission ; Mitochondrial Swelling ; drug effects ; Saponins ; pharmacology ; Time Factors ; Triterpenes ; pharmacology ; Uterine Cervical Neoplasms ; pathology ; ultrastructure