title: Tanshinone IIA inhibits Hep-J5 cells by increasing calreticulin, caspase 12 and GADD153 protein expression.
abstract: Tanshinone IIA (Tan-IIA) is extracted from Danshen, Salviae miltiorrhizae Radix, which has been widely adopted in traditional herbal medicine to treat cardiovascular and hepatic diseases. Tan-IIA induces apoptosis and inhibits proliferation in human hepatocellular carcinoma (HCC) cells. However, the molecular mechanisms of Tan-IIA on human HCC cells are not understood clearly. In the present study, the cytotoxicity of Tan-IIA as well as its molecular mechanisms in human HCC J5 cells was investigated. The cytotoxicity was assayed by MTT. The protein expression of p53, p21, Bax, Bcl-2, Cdc25c, Cdc2, calreticulin, caspase 12, GADD153, caspase 3 and beta-actin in J5 cells were determined by Western blotting. The cell cycles were analyzed by FACS. The protein expression of caspase 12, GADD1533 and caspase 3 were detected by immunocytochemical staining. The results showed that Tan-IIA inhibited J5 cells in a dose- and time-dependent manner. The protein expression of p53, p21, Bax, calreticulin, caspase 12, caspase 3 and GADD153 were increased, but Bcl-2, Cdc25c and Cdc2 were decreased in J5 cells. In addition, the results also showed that Tan-IIA arrested J5 cells in the G2/M phase. Immunocytochemistry staining showed that J5 cells treated with Tan-IIA up-regulated the protein expression of caspase 12, 3 and GADD153. Taken together, the findings suggest that Tan-IIA inhibits and induces apoptosis in J5 cells through novel molecular targets, calreticulin, caspase 12 and GADD153.
authors.affiliation: Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan, ROC.
authors.forename: Chun-Yuan
authors.initials: CY
authors.lastname: Cheng
authors.forename: Chin-Cheng
authors.initials: CC
authors.lastname: Su
mesh_terms: D045784:Abietanes; D000590:Amino Acid Chloromethyl Ketones / Q000378:metabolism; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects; D037282:Calreticulin / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy*; D053454:Caspase 12 / Q000378:metabolism*; D061945:Caspase Inhibitors; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000494:pharmacology / Q000627:therapeutic use; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy*; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D010616:Phenanthrenes / Q000494:pharmacology* / Q000627:therapeutic use; D051742:Transcription Factor CHOP / Q000378:metabolism*
chemical_list: D045784:Abietanes; D000590:Amino Acid Chloromethyl Ketones; D000972:Antineoplastic Agents, Phytogenic; D037282:Calreticulin; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D004365:Drugs, Chinese Herbal; D010616:Phenanthrenes; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C021751:tanshinone; D051742:Transcription Factor CHOP; D053454:Caspase 12