OBJECTIVETo investigate whether paclitaxel (Taxel) can efficiently induce apoptosis of ACC-2 or not, and to study the relation of apoptosis and arrest of cell mitosis.

METHODSPaclitaxel-induced arrest of cell mitosis and apoptosis of ACC-2 cells in various concentration and different treat time were determined using transmission electron microscope (TEM), fluorescence microscope, flow-cytometry and DNA agarose gel electrophoresis technique.

RESULTSUnder fluorescence microscope, apoptotic cells were green with irregular clumping of nucleus chromatin, or even nuclear chromatin segregation. The typical ultra-structural changes of apoptosis observed by TEM were cell compaction, margination of nuclear chromatin, condensation of cytoplasm, protuberances and apoptotic body. "DNA Ladder" was absent in agarose gel electrophoresis of DNA extracted from culture of ACC-2 cells and paclitaxel-induced ACC-2 cells. "Sub-G(1)" phase peak of ACC-2 cells induced by 50 nmol/L paclitaxel in 48 h and 72 h was 17.13% and 16.26%, respectively. The percentage of G(2)/M phase increased in accordance with raise of the paclitaxel concentration and prolongation of treatment. The typical ultra-structural changes of apoptosis were observed in case that G(2)/M phase was arrested.

CONCLUSIONSPaclitaxel could induce apoptosis of ACC-2 cells. Arrest of G(2)/M phase might induce apoptosis of ACC-2 cells.

Apoptosis ; drug effects ; Carcinoma, Adenoid Cystic ; pathology ; ultrastructure ; Dose-Response Relationship, Drug ; G2 Phase ; drug effects ; Humans ; Mitosis ; drug effects ; Paclitaxel ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo investigating the relation between the expression of P-glycoprotein and Glutathione transferase-pi and the chemoresistance.

METHODSThe expressions of these two proteins in patients with oral and maxillofacial squamous carcinoma and normal oral tissues were detected by immunohistochemistry.

RESULTSThe positive expression rate of P-gp and GST-pi in oral and maxillofacial malignant tumor was 57.1% and 53.6% respectively, and no expression in normal oral tissues; the expression of GST-pi was relevant to the resistance to cisplatin, while the expression of P-gp was relevant to the resistance to chemotherapeutic drug in general.

CONCLUSIONSThe method of immunohistochemistry combining MTT assay in vitro may become an efficient way to predict the sensitivity to chemotherapeutic drug.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Carcinoma, Squamous Cell ; chemistry ; drug therapy ; Drug Resistance, Neoplasm ; Facial Neoplasms ; chemistry ; drug therapy ; Formazans ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Maxillary Neoplasms ; chemistry ; drug therapy ; Mouth Neoplasms ; chemistry ; drug therapy ; Tetrazolium Salts

Background and Objectives: Cancer stem cells (CSCs) with tumorigenic potential are reported as the crucial factors of hepatocellular carcinoma (HCC) recurrence and therapy resistance. Bone mesenchymal stem cells (BMSCs) are documented to play an important role in the protection of hepatocytes. Bie Jia Jian pill (BJJP), a Traditional Chinese Medicine, has been used to treat liver fibrosis and liver cancer. This study aimed to explore the potential role of combined use of BJJP with BMSCs in HCC cell lines. Methods: and Results: Flow cytometry was used to identify BMSCs isolated from BALB/c mice and CSCs enriched from Huh7 cells by measuring CD24, CD133, CD44, CD73, CD105, CD166, CD29, CD14 and CD34. Differentiation potential of BMSCs was also determined. Cell viability and proliferation ability of CSCs were determined by CCK-8 assay and clone formation assay. The expressions of CSCs biomarkers and Wnt/β-catenin signal pathway related proteins were determined by PCR and western blot. TOP-Flash/FOP-Flash luciferase assay was applied to measure the activity of β-catenin/TCF. Compared with untreated CSCs, BJJP or BMSCs treatment alone on CSCs lead to increased miR-140 expression and cell apoptosis, as well as decreased expressions of CD24, CD133, EpCAM and cell viability.Downregualted expressions of Wnt/β-catenin signal pathway related proteins, Wnt3a and β-catenin were found in response to BJJP or BMSCs treatment alone. The combination of BJJP＋BMSCs treatment on CSCs could further enhance the suppressive effect on CSCs. Down-regulation of miR-140 in CSCs partially blocked the effects of BMSCs or BMSCs＋BJJP on the expressions of Wnt3a and β-catenin as well as the cell viability and apoptosis of CSCs.Reversed expression pattern was found in CSCs transfected with miR-140 overexpression. Conclusions Taken together, we demonstrate that BJJP＋BMSCs together could further enhance the suppressive effect on CSCs through regulating miR-140 and suppressing Wnt/β-catenin signal pathway. This study demonstrated the potential of BJJP＋BMSCs in therapeutic treatment of HCC.

Background and Objectives: Cancer stem cells (CSCs) with tumorigenic potential are reported as the crucial factors of hepatocellular carcinoma (HCC) recurrence and therapy resistance. Bone mesenchymal stem cells (BMSCs) are documented to play an important role in the protection of hepatocytes. Bie Jia Jian pill (BJJP), a Traditional Chinese Medicine, has been used to treat liver fibrosis and liver cancer. This study aimed to explore the potential role of combined use of BJJP with BMSCs in HCC cell lines. Methods: and Results: Flow cytometry was used to identify BMSCs isolated from BALB/c mice and CSCs enriched from Huh7 cells by measuring CD24, CD133, CD44, CD73, CD105, CD166, CD29, CD14 and CD34. Differentiation potential of BMSCs was also determined. Cell viability and proliferation ability of CSCs were determined by CCK-8 assay and clone formation assay. The expressions of CSCs biomarkers and Wnt/β-catenin signal pathway related proteins were determined by PCR and western blot. TOP-Flash/FOP-Flash luciferase assay was applied to measure the activity of β-catenin/TCF. Compared with untreated CSCs, BJJP or BMSCs treatment alone on CSCs lead to increased miR-140 expression and cell apoptosis, as well as decreased expressions of CD24, CD133, EpCAM and cell viability.Downregualted expressions of Wnt/β-catenin signal pathway related proteins, Wnt3a and β-catenin were found in response to BJJP or BMSCs treatment alone. The combination of BJJP＋BMSCs treatment on CSCs could further enhance the suppressive effect on CSCs. Down-regulation of miR-140 in CSCs partially blocked the effects of BMSCs or BMSCs＋BJJP on the expressions of Wnt3a and β-catenin as well as the cell viability and apoptosis of CSCs.Reversed expression pattern was found in CSCs transfected with miR-140 overexpression. Conclusions Taken together, we demonstrate that BJJP＋BMSCs together could further enhance the suppressive effect on CSCs through regulating miR-140 and suppressing Wnt/β-catenin signal pathway. This study demonstrated the potential of BJJP＋BMSCs in therapeutic treatment of HCC.