Mechanism of Platycarya strobilacea Sieb. et Zucc extract-induced methuosis in human nasopharyngeal carcinoma CNE1 and CNE2 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of Platycarya strobilacea Sieb. et Zucc (PSZ) extract on methuosis of human nasopharyngeal carcinoma CNE1 and CNE2 cells and explore the underlying mechanism.</p><p><b>METHODS</b>CNE1 and CNE2 cells were treated with 1 mg/mL PSZ extract and the expressions of Rac1 mRNA and Rac1 protein were detected using RT-qPCR and Western blotting, respectively. Results CNE1 and CNE2 cells showed obvious morphological changes typical of methuosis following treatment with PSZ extract characterized by cell merging, accumulation of large cytoplasmic vacuoles, and membrane rupture without obvious changes in the nuclei. PSZ treatment resulted in up-regulated Rac1 mRNA and Rac1 protein expressions in the cells. Application of EHT 1864 obviously blocked the effect of PSZ extract in inducing methuosis in CNE1 and CNE2 cells.</p><p><b>CONCLUSION</b>PSZ extract can induce methuosis in CNE1 and CNE2 cells by inducing the overexpression of Rac1.</p>

Mechanism of thioridazine-induced apoptosis of human colorectal cancer SW480 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of thioridazine on the proliferation and apoptosis of human colorectal cancer SW480 cells.</p><p><b>METHODS</b>SW480 cells were treated with different concentrations of thioridazine, and MTT assay was used to evaluate the cell inhibition rate. Hoechst 33342 staining was performed to demonstrate the cell morphology changes. Flow cytometry was used to determine the cell apoptosis and cell cycle changes. RT-qPCR was used to detect PDCD4, c-MYC, BCL2, CCND1, CASPASE3, PARP1, CDK4 and EIF4A mRNA expressions, and Western blotting was employed to assay AKT, p-AKT, and PDCD4 protein expression levels.</p><p><b>RESULTS</b>MTT results showed that thioridazine inhibits the proliferation of SW480 cells. SW480 cells treated with thioridazine presented with such typical features of apoptosis of karyopyknosis, chromatin condensation and nuclear fragmentation. Flow cytometry showed that thioridazine was a cell cycle-specific drug and caused cell cycle arrest at G(1)/G(0) phase and an increased cell apoptosis rate. Thioridazine treatment of the cells resulted in up-regulated PDCD4 mRNA expression and down-regulated mRNA expressions of CCND1, CDK4, c-MYC, BCL2, CASPASE3, PARP1 and EIF4A, increased PDCD4 protein expression and reduced p-AKT protein expression.</p><p><b>CONCLUSION</b>Thioridazine inhibits the proliferation and induces apoptosis of SW480 cells by up-regulating PDCD4 and inhibiting PI3K/Akt pathway.</p>

Extract of Zuojin Pill (characters: see text) induces apoptosis of SGC-7901 cells via mitochondria-dependent pathway / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of water extract of Zuojin Pill ([characters: see text], ZJP) on inhibiting the growth of human gastric cancer cell line SGC-7901 and its potential mechanism.</p><p><b>METHODS</b>Effects of ZJP on SGC-7901 cells growth were determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, cell apoptosis and cell cycle were determined by flow cytometry, and apoptosis induction was detected by means of DNA gel electrophoresis. The cellular mechanism of drug-induced cell death was unraveled by assaying oxidative injury level of SGC-7901 cell, mitochondrial membrane potentials, expression of apoptosis-related genes, such as B cell lymphoma/lewkmia-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cleaved caspase-3 and caspase-9.</p><p><b>RESULTS</b>ZJP exerted evident inhibitory effect on SGC-7901 cells by activating production of reactive oxygen species and elevating Bax/Bcl-2 ratio in SGC-7901 cells, leading to attenuation of mitochondrial membrane potential and DNA fragmentation.</p><p><b>CONCLUSIONS</b>ZJP inhibits the cancer cell growth via activating mitochondria-dependent apoptosis pathway. ZJP can potentially serve as an antitumor agent.</p>

STAT3 correlates with stem cell-related transcription factors in cervical cancer / 华中科技大学学报（医学）（英德文版）

Cancer stem cells (CSCs) are considered responsible for the high recurrence rate in cervical carcinoma. It has been demonstrated that the signal transducer and activator of transcription 3 (STAT3) is involved in the oncogenesis and takes part in mediating the effects of maintaining stem cell phenotype and pluripotency by regulating the expression of stem cell-related transcription factors. However, the correlation between STAT3 and stem cell-related transcription factors in cervical cancer has not been elucidated. In this study, we established overexpressing plasmid (GV316-STAT3) and siRNA-STAT3 for transfecting Siha cells. Cells negative or positive for Nanog, Oct4, or Sox2 were selected by flow cytometry. Proliferation and differentiation rate of Siha cells was determined by detecting the efficiency of tumor sphere formation. The expression of Nanog, Oct4 and Sox2 (cancer stem cell markers) and STAT3 was detected by quantitative real-time PCR and immunoblotting for Siha cells and by immunohistochemistry (IHC) for cervical tissues, respectively. The results showed that Nanog+, Oct4+, and Sox2+ Siha-STAT3 over-expressing cells displayed the typical non-adherent spheres. The sphere formation efficiency was significantly different between Siha-STAT3 overexpressing cells and siRNA-STAT3 cells (P<0.05). Meanwhile, the expression levels of Oct4, Nanog and Sox2 mRNA and protein were significantly higher in Siha-STAT3 overexprssing cells than in siRNA-STAT3 cells (P<0.05). In addition, the positive rate of STAT3, Nanog, Oct4 and Sox2 in cervical cancer tissues was higher than that in chronic cervicitis group (P<0.05). There was a significantly positive relationship between STAT3 and Nanog or Oct4 or Sox2 expression (all P<0.001). These results suggested that Oct4+, Sox2+, and Nanog+ cell population possesses stem cell properties in cervical cancer, which may contribute to cervical carcinogenesis and be regulated by STAT3.

Effect of curcumin on radiosensitization of CNE-2 cells and its mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.</p><p><b>METHOD</b>The effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.</p><p><b>RESULT</b>After 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.</p><p><b>CONCLUSION</b>Cur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.</p>

Proportion of Coptidis rhizoma and Evodiae fructus in the compound preparation: its effect in inducing apoptosis of SGC-7901 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of water extracts of Coptidis Rhizoma and Evodiae Fructus (CREF) on the proliferation and apoptosis of human gastric carcinoma cells (SGC-7901) and determine the optimal proportion of Coptidis rhizoma to Evodiae fructus.</p><p><b>METHODS</b>The growth inhibition of SGC-7901 cells treated with the water extracts of CREF of varying proportions was tested with MTT assay. The cell apoptotic rate and mitochondrial membrane potential were analyzed with flow cytometry.</p><p><b>RESULTS</b>The water extract of CREF with Coptidis Rhizoma: Evodiae Fructus proportions at 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1 all significantly inhibited the growth of SGC-7901 cells after a 24-h or 48-h treatment (P<0.05). The growth inhibition and cell death ratio both exhibited a dose-dependent pattern of Coptidis Rhizoma. Flow cytometry analysis showed that, after treatment of the cells with CREF at the proportions of 1:6, 2:5, 3:4, 4:3, 5:2, and 6:1, the apoptotic rate were (8.50 ∓ 1.59)%, (9.90 ∓ 1.01)%, (17.15∓1.68)%, (21.55 ∓ 1.97)%, (34.10 ∓ 1.06)% and (34.40 ∓ 1.02)%, respectively, all significantly higher than that in the control group [(1.69 ∓ 1.91)%, P<0.05]. JC-1 Kit staining showed that mitochondrial membrane potential of SGC-7901 cells was decreased and the ratio of green to red fluorescence increased significantly after incubation with CREF.</p><p><b>CONCLUSION</b>CREF can inhibit the growth and induce apoptosis of SGC-7901 cells, and the strongest effect is achieved at the optimal proportion of Coptidis Rhizoma and Evodiae Fructus at 6:1.</p>

Establishment of a nasopharyngeal carcinoma cell line CNE1 stably overexpressing ebv-miR-BART7 mediated by lentivirus / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a nasopharyngeal carcinoma (NPC) cell line CNE1-pLVTHM/BART7 with stable ebv-miR-BART7 overexpression.</p><p><b>METHODS</b>The recombinant lentivirus pLVTHM/BART7 expression plasmid was packaged into mature lentivirus by 293FT cells and used to infect CNE1 cells. Flow cytometry was employed for sorting the GFP(+) cells. The efficiency of ebv-miR-BART7 overexpression was determined using qRT-PCR.</p><p><b>RESULTS</b>The recombinant lentivirus plasmid pLVTHM/BART7 was successfully constructed and verified by PCR and sequencing. The expression of ebv-miR-BART7 in CNE1 cells infected with the lentivirus pLVTHM/BART7 was significantly increased as compared with the negative control and the blank control cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus vector pLVTHM/BART7 results in high and stable expression of ebv-miR-BART7 in infected CNE1 cells, which provides a useful cell model for further studies of the role of ebv-miR-BART7 in nasopharyngeal carcinoma.</p>

Analysis of the relative distribution and gene variation of HPV16 transforming gene at different stages of cervical lesions / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to analyze the relative distribution and gene variation of HPV16 transforming gene E6, E7 and E5 at different stages of cervical lesions.</p><p><b>METHODS</b>DNA was extracted from tissue samples of 200 patients with cervical lesions, including 124 cervical cancer, 17 CIN grade I and II, 23 CIN grade III and 36 cervicitis. Then HPV16 E6, E7 and E5 genes were amplified, and part of the E6 and E7 PCR products were sequenced using the HPV16 E6 and E7 specific primers.</p><p><b>RESULTS</b>The positive rate of E6 gene in cervicitis, CINI and CINII, CINIII and cervical cancer was 25.0%, 29.4%, 60.9% and 76.6%, respectively. The positive rate of E7 gene was 16.7%, 41.2%, 43.5% and 61.3%, respectively. The positive rate of E5 gene was 5.6%, 5.9%, 30.4% and 40.3%, respectively. HPV16 E6 gene mutations in Nt 178 were found in 47 case from 80 cervical cancer samples, resulting in amino acid change of Asp to Glu. The mutation rate was 58.8%.Otherwise the mutation rate of E6 178 in cervicitis and CIN I approximately III samples was 25.0% and 31.8%. E7 mutations were found in Nt 647 in 21 cervical samples from 30 cervical cancer samples, resulting in amino acid change of Asn to Ser. The mutation rate was 70.0%. The mutation rate of E6 647 in cervicitis and CINI approximately III samples was 35.0% and 40.9%, respectively.</p><p><b>CONCLUSION</b>The positive rate of E6 and E7 increase gradually from cervicitis, CINI and CINII, CINIII to cervical cancer. The rate of E5 is relatively lower than that of E6 and E7 gene in cervical tissue samples. These results show that E6 and E7 gene are highly associated with the progress of cervical cancer and E5 genes are lost in the development of cervical cancer. High frequency mutations of HPV16 E6 and E7 gene in E6 178, E7 647 have been found in cervical cancer samples in Hubei province, China. These results approved that the HPV16 variants prevalent in this area are different from the European and African variants.</p>

Proteomic analysis of chronic restraint stress-induced Gan (肝)-stagnancy syndrome in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To analyze the proteomic characteristics of Gan (肝)-stagnancy syndrome (GSS) by seeking the differential protein in blood and tissues of GSS model rats.</p><p><b>METHODS</b>GSS model rats were established by chronic restraint stress, keeping rats in restrain chamber for 6 h every day for 21 successive days. Their blood and liver samples were collected at the end of experiment for differential protein detection with methods of isoelectrofocusing and polyacrylamide SDS-PAGE, silver staining, and scanning. The gel images were analyzed with Imagemaster 2D Elite software, and the excavated differential protein spots were identified with matrix assistant laser resolving TOF mass spectrometry, Western blot, ELISA, and RT-PCR, respectively.</p><p><b>RESULTS</b>A method for isolating the protein in blood serum and tissues by two-dimensional gel electrophoresis was established and optimized. Six serum proteins and three liver proteins that differentially expressed were identified. The down-regulated differential proteins in serum of GSS model rats were serum albumin precursor, beta 1 globin, antibody against muscle acetylcholine receptor, Ig lambda-2 C region, and transthyretin (TTR), and those in liver tissue were aryl sulfotransferase, enoyl-CoA hydratase, and TTR. TTR down-regulation was found in both serum and liver. Preliminary biological information analysis showed that these differential proteins involved in immune, neuroendocrine, nutrition, and substance metabolism.</p><p><b>CONCLUSION</b>Proteomic analysis of differential proteins showed that TTR, aryl sulfotransferase, and enoyl-CoA hydratase expressions are downregulated in the GSS model rats, suggesting that the susceptibility of cancer could be enhanced by chronic stress.</p>

Guling tablets promotes the proliferation and mineralization and regulates OPG/RANKL/M-CSF expression of osteoblasts via the p38 MAPK pathway in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Gulingpian (GLP, or Guling tablets) on the proliferation and mineralization of MG-63 cells and the involvement of p38 MAPK pathway in such processes.</p><p><b>METHODS</b>MG-63 cells were cultured in the presence of p38 inhibitor SB203580 and or serum from rats fed with normal saline or GLP. The expression of the OPG, RANKL and M-CSF mRNA, cell proliferation and formation of mineralized bone nodule, the total protein of p38 and phosphorylated p38, were observed or quantitatively measured.</p><p><b>RESULTS</b>Treatment with GLP serum increased the level of p38 phosphorylation but did not affect the total p38 expression. GLP serum significantly promoted MG-63 cell proliferation and differentiation, resulting also in up-regulated OPG and down-regulated RANKL mRNA expressions without obvious alteration in M-CSF expression. These effects were blocked by the specific p38 inhibitor SB203580.</p><p><b>CONCLUSION</b>GLP promotes the proliferation and differentiation of osteoblasts and regulates OPG/RANKL expressions in vitro, which are mediated probably via the p38 MAPK pathway.</p>