Relationship between apoptosis,proliferation and expression,mutation of related genes in breast cancer

AIM:To study the relationship between apoptosis, proliferation and expression,mutation of related genes in breast cancer.METHODS:Methods of TUNEL, immunohistochemical S-P and PCR-SSCP were used respectively to study apoptotic index (AI), mitotic index(MI), expression of Bcl-2,p53,c-erbB-2,PCNA,Ki67,TopoⅡ and mutation of p53 in 54 cases of breast cancer.RESULTS:AI and MI were 9.40±3.78 and 5.96±2.36, respectively. There was a significant direct correlation between them(r=0.46.P＜0.01).High expression of Bcl-2,PCNA,Ki67,TopoⅡ coincided with high AI,MI(P＜0.01). High expression of p53,c-erbB-2 and mutation of p53 coincided with high MI(P＜0.01). Type of p53 mutation coincided with AI(P＜0.05).CONCLUSION:Disturbance of gene control between apoptosis and proliferation is related with expression,mutation of related genes in breast cancer.     

Parthenolide enhances the apoptosis induced by 4-hydroxyphenyl-retinamide in human hepatoma cells

AIM:To detect the signal pathway of apoptosis induced by 4-hydroxyphenyl-retinamide (4-HPR) and the biological effect of parthenolide-induced apoptosis. METHODS:TUNEL staining, FCM analysis, electrophoretic mobile shift assay (EMSA) were used to determine the actual effects and its mechanism of parthenolide on the 4-HPR-induced apoptosis in human hepatoma Hep-3B and SK-Hep-1 cells. RESULTS:The results of TUNEL and PI staining showed that parthenolide selectively enhanced 4-HPR-induced apoptosis in Hep-3B and SK-Hep-1 cells. Subsequent observations using EMSA assay indicated that parthenolide effectively inhibited NF-κB activation during fenretinide-induced apoptosis. CONCLUSION:These findings indicate that parthenolide suppresses 4-HPR-induced apoptosis via inhibition of NF-κB activation and that NF-κB activation during fenretinide-induced apoptosis might have an anti-apoptotic effect.     

Effects of erythromycin and its derivatives on the proliferation of T lymphocyte in vitro

AIM: To study the effects of erythromycin and its derivatives on the proliferation and the apoptosis of T lymphocytes and to evaluate the anti-inflammatory mechanism of erythromycin derivatives. METHODS: The effects of four erythromycin derivatives without antibacterial action on the proliferation T lymphocytes were evaluated by MTT. The effects of the erythromycin derivatives on apoptosis in T lymphocytes were observed by flow cytometry and TUNEL staining. RESULTS: The proliferation of T lymphocytes were inhibited by erythromycin derivative-Ⅰ and erythromycin, the IC50 was (425.2±32.1) μmol/L and (606.3±35.4) μmol/L, respectively. The IC50 value of erythromycin derivative-Ⅰ was lower than that of EM (P<0.01). Erythromycin derivative-Ⅰ (3-30 mg/L) and EM (30-100 mg/L) induced T lymphocytes apoptosis in a concentration-related manner by flow cytometry and TUNEL, while erythromycin derivative-Ⅰ induced cells to die at concentration of 100 mg/L. CONCLUSION: The anti-inflammatory activity of erythromycin derivatives may be due partly to their effects on the proliferation and apoptosis of T lymphocytes.     

Effect of HIPK2 gene on viability,apoptosis and JAK2/STAT3 signaling pathway in NRK-52E renal tubular epithelial cells induced by hypoxia and reoxygenation

AIM: To investigate the effect of homeodomain-interacting protein kinase 2 (HIPK2) on the viabi-lity, apoptosis and JAK2/STAT3 signaling pathway in NRK-52E renal tubular epithelial cells induced by hypoxia and reoxygenation (H/R). METHODS: HIPK2 small interfering RNA (siRNA) was transfected into NRK-52E cells by Lipofectamine^TM 2000, and normal control group (control group) and negative control group (HIPK2-NC group) were set up. After H/R, the cell viability was measured by CCK-8 assay, the apoptotic rate and Ca²⁺ fluorescence intensity were analyzed by flow cytometry, and the protein levels of Ki67, cleaved caspase-3, caspase-12, Bcl-2, Bax, p-JAK2 and p-STAT3 were determined by Western blot. RESULTS: Compared with control group, the protein expression of HIPK2 in the NRK-52E cells was significantly decreased after transfection with HIPK2 siRNA (P<0.05). Compared with control group, the cell viability and the protein expression of Ki67 and Bcl-2 in H/R group were also significantly decreased, and the apoptotic rate, the Ca²⁺ fluorescence intensity and the protein levels of cleaved caspase-3, caspase-12, Bax, p-JAK2 and p-STAT3 were significantly increased (P<0.05). Compared with H/R group, the cell viability and the protein expression of Ki67 and Bcl-2 in HIPK2-siRNA+H/R group were significantly increased, while the apoptotic rate, the Ca²⁺ fluorescence intensity and the protein levels of cleaved caspase-3, caspase-12, Bax, p-JAK2 and p-STAT3 were significantly decreased (P<0.05). CONCLUSION: Inhibition of HIPK2 gene expression promotes H/R-induced growth of NRK-52E renal tubular epithelial cells, and reduces the apoptosis. The mechanism is related to down-regulating the JAK2/STAT3 signaling pathway.     

Effect of captopril on AGS nude mouse model of gastric cancer

AIM: To observe the effect of captopril on the genesis and development of gastric cancer, and to explore its clinical treatment feasibility for gastric cancer. METHODS: The human gastric cancer cell line AGS was used to establish a tumor model in nude mice, and the model mice were randomly divided into 3 groups: positive control (5-fluorouracil) group, normal control (saline) group and experimental (captopril) group. After intraperitoneal injection or intragastric administration of the drugs, the tumor growth curve was determined, and the tumor tissues were also sampled to detect the expression of Ki-67, STAT3, Bax and Bcl-2 by real-time quantitative PCR and immunohistochemistry. The apoptosis was detected by TUNEL+DAPI staining. RESULTS: The tumor growth curve showed that the tumor model in the nude mice was successfully established. The tumor volumes among groups showed significantly different after 14 d growth. The increase in the tumor volume in normal control group was significantly faster than that in the other two groups, and that in positive control group was the slowest. The expression of Bax in captopril group increased, and the expression of STAT3, Ki-67 and Bcl-2 was reduced as compared with normal control group and positive control group. Compared with normal control group, the apoptotic rate increased significantly, and the protein expression of p-STAT3 and STAT3 decreased obviously in positive control group and captopril group. CONCLUSION: With better feasibility, angiotensin-converting enzyme inhibitor captopril has a significant effect on treating gastric cancer in the AGS nude mouse model by regulating the expression of STAT3, Bax, Bcl-2 and Ki-67 to accelerate the apoptosis of cancer cells, thus inhibiting tumor growth.     

Preliminary separation and identification of quiescent and activated neural stem cells in mouse embryonic cerebral cortex

AIM To isolate and identify quiescent and activated neural stem cells from mouse embryonic cerebral cortex. METHODS Two cell clusters derived from mouse cerebral cortex on embryonic day 14.5 were separated by flow cytometry. The expression of stem cell marker Pax6 and proliferation marker Ki67 was examined by immunofluorescence. The mRNA expression of stem cell marker genes Pax6, Oct4, Sox2 and Nanog were detected by RT-qPCR. Cell cycle was analyzed by flow cytometry and RT-qPCR. Proliferation ability was investigated by in vitro cell culture. RESULTS In both 2 groups, the cells expressed Pax6. Immunofluorescence staining of Ki67 in the big cell group was positive, while that in small cell group was negative. Cell cycle assay showed that the proportion of G₀/G₁ phase in the small cells was higher than that in the big cells, the G₂/M phase proportion was 0, and the expression of cyclin A and cyclin B was lower than that in the big cells (P<0.05). When cultured in vitro, the number of microspheres formed by the small cells was smaller and the formation speed was slower than those of the big cells. After digestion of microspheres, Pax6 and Ki67 staining of both big and small cells was positive, and the positive rates were not different (P>0.05), indicating that the quiescent neural stem cells were activated. CONCLUSION The 2 cell clusters are quiescent and activated state of neural stem cells. The activated stem cells have strong abilities of self-renewing and proliferation, while these abilities of quiescent stem cells are poor. The quiescent stem cells can translate into activated ones when cultured in vitro for a period.     

Effects of tangeretin on growth and invasion of human non-small-cell lung cancer cells

AIM: To study the influences of tangeretin (TGN) on the growth and invasion of non-small-cell lung cancer (NSCLC) cells, and to explore the molecular mechanisms. METHODS: The A549 cells were treated with different concentrations of TGN in vitro. The relative cell activity was determined by MTT assay. The apoptotic rate was analyzed by flow cytometry with Annexin V-FITC/PI staining. The number of the invasive cells was measured by Transwell assay. The mRNA expression of MMP-2 and MMP-9 was detected by RT-PCR, and the protein levels of Ki67, Cyt C, caspase-3, cleaved caspase-3, MMP-2, MMP-9, Akt, p-Akt and p-PI3K were determined by Western blotting analysis. RESULTS: TGN inhibited the proliferation of A549 cells in a dose-dependent manner (P<0.05) along with the low expression level of proliferation biomarker Ki67. TGN up-regulated the protein levels of Cyt C, caspase-3 and cleaved caspase-3 (P<0.01) and promoted the apoptosis of A549 cells in a dose-dependent manner. Moreover, TGN down-regulated the invasion-related molecules MMP-2 and MMP-9 at the mRNA and protein levels, and the number of invasive cells reduced with the increase in the concentration of TGN. The protein levels of p-Akt and p-PI3K in the A549 cells was reduced (P<0.05), and no difference of the cell viability in the cells treated with different concentrations of TGN was observed after blocking PI3K/Akt signaling pathway using LY294002. CONCLUSION: TGN inhibits the growth and invasion of A549 cells and promotes the cell apoptosis by potentially inhibiting PI3K/Akt signaling pathway activation. Therefore, this study will provide a new target for the prevention and control of NSCLC.     

Effect of hPTTG1 siRNA on proliferation and apoptosis of ovarian cancer cells

AIM: To observe the proliferation and apoptosis of ovarian cancer cells by silencing the expression of human pituitary tumor-transforming gene 1 ( hPTTG1 ) using RNA interference technique.METHODS: The chemically synthesized siRNA targeting hPTTG1 was transfected into ovarian cancer cell line A2780 in vitro. The expression levels of hPTTG1 and c-myc were examined by RT-PCR and Western blotting. Cell proliferation was measured by MTT colorimetric assay and -TdR incorporation test. Cell apoptosis was detected by flow cytometry with annexin V/PI and TUNEL labeling.RESULTS: The expression of hPTTG1 at mRNA and protein levels was inhibited after transfection of hPTTG1 siRNA. The inhibitory efficiency was 70.5%±3.9% and 63.8%±4.5%, respectively. The absorbance began to decrease 24 h after transfection of hPTTG1 siRNA,and the highest inhibitory rate was 42.9%±5.2% at 48 h post-transfection. Radioactive incorporation of -TdR in hPTTG1 siRNA group was lower than that in normal and negative groups. The survival rate declined while the apoptotic rate and necrotic rate increased in hPTTG1 siRNA group. Apoptotic index in hPTTG1 siRNA group was higher than that in normal and negative groups. The expression of c-myc at mRNA and protein levels was down-regulated.CONCLUSION: Cell proliferation is inhibited and cell apoptosis is induced by hPTTG1 siRNA through down-regulating the expression of c-myc. hPTTG1 can be regarded as a candidate gene for ovarian cancer gene therapy.     

Expression of VEGFR-3, CD31 and their correlation with metastasis in ovarian epithelial tumors

AIM: To investigate the expression of vascular endothelial growth factor receptor-3 (VEGFR-3) and CD31 in relation to metastasis in ovarian epithelial tumors. METHODS: VEGFR-3 and CD31 expression were examined by immunohistochemical methods, VEGFR-3 positive microlymphatic count (MLC) and microvessel density (MVD) were assessed by the image analysis. RESULTS: Both MLC and MVD in ovarian epithelial carcinomas were higher than those in benign tumors(MLC, P＜0.05; MVD, P＜0.01). In ovarian epithelial carcinomas, MLC and MVD were higher in the cases of clinical stage Ⅲ-Ⅳ and with lymphatic metastasis than those of clinical stage Ⅰ-Ⅱ and without lymphatic metastasis, respectively (P＜0.05 or P＜0.01) . There were no significant differences between MLC, MVD and histologic type, histologic grade(differentiation) in ovarian epithelial carcinomas(P＞0.05). CONCLUSIONS: VEGFR-3 and CD31 expression correlate significantly with metastasis, MLC and MVD might predict peritumoral lymphangiogenesis and angiogenesis, which may be as a biologic marker for metastasis in ovarian epithelial tumors.     

Effect of conjunction matrigel with MFP implantation on the tumorigenesis,proliferation, apoptosis and metastasis of breast cancer cells with different expression of Her2

AIM: To detect the effect of conjunction matrigel with mammary fad pat(MFP)implantation on the tumorigenesis, proliferation, apoptosis and metastasis of Her2 positive and negative breast cancer model. METHODS: The Her2 positive BT 474 and Her2 negative MDA-MB 231 breast cancer cells were injected into MFP of nude mice with or without matrigel to establish breast cancer model. The tumor volume was measured every 3 d. Followed up for 30 d after implantation, the nude mice were killed and the tumors and associated organs were dissected for pathological sectioning and staining with hematoxylin and eosin. The time of tumor formation and the tumorigenesis were determined after implantation. The tumor volume and metastasis rate were calculated and compared with each other. The proliferation and apoptosis of Her2 positive and negative tumors were also determined. RESULTS: Matrigel and MFP implantation technology shortened the time of tumorigenesis significantly(P<0.01). The tumorigenesis rate of BT 474 and MDA-MB 231 breast cancer cells did not show any different(P>0.05). The metastasis rate of MDA-MB 231 breast cancer cells were improved from 25.0% to 37.5%(P<0.05). CONCLUSION: Matrigel and MFP implantation can be combined to shorten the time of tumor formation by two kinds of breast cancer cells, and improve the metastasis of Her2 negative MDA-MB 231 cells. Using matrigel does not show any effect of proliferation and apoptosis on Her2 positive and negative breast cancer cells.     

Correlation between Helicobacter pylori infection and apoptosis in gingival tissues

AIM:To observe the effects of Helicobacter pylori(Hp) on the apoptosis of human gingival tissue.METHODS:Gingival tissue samples were taken from 30 patients without chronic periodontitis,and Hp was detected by conventional PCR.The apoptosis of the gingivival cells was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to analyze the correlation between Hp infection and apoptosis of the gingival tissues.RESULTS:The Hp positive detections were 12 in the 30 patients without periodontitis,so the positive rate of Hp in the gingival tissue samples was 40%.The gingival tissue showed a large number of apoptotic cells in Hp positive group,and less apoptotic cells in Hp negative group.The apoptotic index in Hp positive group (0.498±0.092) was significantly higher than that in normal group (0.207±0.053)(P<0.05).CONCLUSION:Hp might play a role in the apoptosis of gingival tissues.     

Effects of baicalin on CA46 cell proliferation inhibition and apoptosis induction

AIM: To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms. METHODS: CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTT assay. The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V-FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation. The mRNA expressions of c-myc and bcl-2 were detected by RT-PCR, and the protein expressions of c-Myc, Bcl-2, caspase-3 precursor (procaspase-3) and poly ADP-ribose polymerase (PARP) were detected by Western blotting. RESULTS: Baicalin remarkably inhibited the CA46 cell proliferation, with an IC50 value of 10 μmol/L. Apoptosis was remarkably induced by baicalin in a dose-dependent manner, and its earlier and later stages were detected by annexin V-FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation, respectively. Furthermore, RT-PCR showed that the mRNA expressions of c-myc and bcl-2 in treated CA46 cells decreased in a time-dependent manner. Western blotting showed that the protein expressions of c-Myc, Bcl-2, procaspase-3 and PARP (116 kD) in baicalin treated CA46 cells were down-regulated in a time-dependent manner, while the expression of PARP(85 kD) was up-regulated. CONCLUSION: Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells, which may be related with the down-regulation of c-Myc and Bcl-2 expressions, as well as the up-regulation of caspase-3 activity.     

Effect of 1, 2-propanediol and ethylene glycol on apoptosis of follicles in human ovarian tissue cryopreservation

AIM:To compare the effect of 1, 2-propanediol (PROH) and ethylene glycol (EG) on apoptosis and expressions of P53, Bcl-2 of follicles in human ovarian tissue, in order to offer experimental foundation for selecting the best cryoprotectant.METHODS:Biopsies of ovarian tissue obtained from 12 women were cryopreserved, and ovarian tissue slice from each woman was divided into three groups:fresh control group, ethylene glycol group and 1, 2-propanediol group.The slow-freezing /rapid-thawing protocol was used to freeze and thaw the slice of ovarian cortex.Apoptosis of follicles in fresh and frozen-thawed ovarian cortical tissue was detected by TUNEL experiment, and expressions of P53 and Bcl-2 were detected by immuno-histochemistry.RESULTS:The percentages of apoptosis follicles were 14.58%, 23.08%, 30.43% in fresh control group, PROH group and EG group, respectively, and the percentage of apoptosis follicles in EG group was higher than that in fresh control group (P<0.05), but there was no significant difference between the percentages of TUNEL positive follicles in oocytes of the three groups (P>0.05).The percentages of P53 positive follicles were 13.48%, 25.00% and 33.93%,respectively.There was significant difference between fresh control and EG group (P<0.05), and there was positive correlation between apoptosis and expression of P53 (r=0.7791, P<0.01).The percentages of Bcl-2 positive follicles were 5.29%, 3.65% and 2.41%, respectively, however, no significant difference between three groups was observed (P>0.05).CONCLUSION:The results of this study indicate that 1.5 mol/L PROH is more suitable for cryopreservation of human ovarian tissue rather than 1.5 mol/L EG in slow-freezing /rapid-thawing protocol.The protocol of slow-freezing/ rapid- thawing may preserve oocytes well, but it is not ideal for cryopreservation of granulosa cells.Cryopreservation may influence on apoptosis of follicles in human ovarian tissue.     

Plumbagin enhances TRAIL-induced apoptosis in Kasumi-1 cells

AIM: To investigate the effect of plumbagin and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on the apoptosis of leukemic Kasumi-1 cells. METHODS: Kasumi-1 cells were treated with plumbagin alone, recombinant soluble TRAIL(rsTRAIL) alone or the combination of plumbagin with rsTRAIL to induce apoptosis. The cell proliferation was analyzed by CCK-8 assay. Apoptosis was determined by flow cytometry with AnnexinⅤ/PI double staining and TUNEL staining. The expression of DR4 and DR5 at mRNA level was measured by real-time PCR. The expression of signal transduction proteins, such as DR5, caspase-3, caspase-8, caspasep-9, Bid, Bax and c-FLIP was detected by Western blotting. RESULTS: Both rsTRAIL and plumbagin induced the apoptosis in Kasumi-1 cells, and combination of plumbagin with rsTRAIL enhanced the apoptosis. The ratios of Annexin V-positive Kasumi-1 cells were (27.7±2.9)%, (25.6±3.1)% and (52.1±3.3)% in 100 μg/L rsTRAIL group, 2 μmol/L plumbagin group and the combination group, respectively, and the positive rate in combination group was significantly higher than those in other 2 groups. TUNEL assay demonstrated that the number of apoptotic cells in combination group was higher than that in the cells treated with rsTRAIL or plumbagin alone. Plumbagin up-regulated the expression of DR5 at mRNA level in Kasumi-1 cells, and up-regulation of DR5, activation of caspase-8 and down-regulation of c-FLIP at protein level were detected in the cells treated with plumbagin alone and the combination of plumbagin with rsTRAIL. CONCLUSION: Plumbagin enhances TRAIL-induced apoptosis in Kasumi-1 cells by up-regulating DR5, activating caspase-8 and down-regulating c-FLIP.     

Apoptosis and Bcl-2/Bax expression in the early follicles of reproductive women

AIM: To investigate the apoptosis and Bcl-2/Bax expression in the early follicles of women at reproductive age. METHODS: 12 ovarian specimens were collected from reproductive women (aged 23-38 years) undergoing gynaecological operation. Histopathological examination of these specimens was performed to confirm its' morphological normalities. Using TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay and immunohistochemistry method, cell apoptosis and Bcl-2/Bax expression were examined in the early follicles including mainly primordial, intermediary and primary follicles. RESULTS: 18.75% of the oocytes were found TUNEL positive in the early follicles, but no granulosa cells in these follicles were found TUNEL positive. Bax expression was detected in 76.07% of the oocytes in the early follicles, but Bcl-2 expression was negative in these oocytes. In addition, Bcl-2/Bax expression were not present in the granulosa cells in early follicles. CONCLUSION: The oocyte apoptosis occurs in the early follicles of reproductive woman, and pro-apoptotic protein Bax may play a role in regulating this process. It suggests that Bax mediated oocyte apoptosis may be the molecular mechanism of the early follicle atresia in the ovaries of reproductive woman.     

The anticancer activity of genistein on implanted tumor of human primary gastric carcinoma cells in nude mice

AIM: To investigate the apoptosis of implanted tumor of primary human gastric cancer cells in nude mice induced by genistein and the relation between this apoptosis and expression of bcl-2 and bax.METHODS: Establishing a transplanted tumor model by injecting human primary gastric cancer cells into subcutaneous tissue of nude mice.The different doses of genistein (0.5mg/kg,1mg/kg and 1.5 mg/kg ) were directly injected beside tumor body respectively,for six times at an interval of two days.Then changes of tumor volume were measured continuously and tumor inhibition rate of each group was calculated.We observed the morphologic alteration by electron microscope,measured the apoptotic rate by TUNEL staining method,detected the expression of apoptosis-regulated gene bcl-2 and bax by immunohistochemical staining and RT-PCR.RESULTS: Genistein could significantly inhibit carcinoma growth when it was injected near the carcinoma.Genistein induced implanted tumors cells to undergo apoptosis with apoptotic characteristics by transmission electron microscope.The apoptosis index of above three groups was increased progressively.Positive rate of Bcl-2 protein of above three groups was decreased progressively and positive rate of Bax protein of above three groups was increased progressively by immunohistochemical staining.The density of bcl-2 mRNA decreased progressively and the density of bax mRNA increased progressively with elongation of time by RT-PCR.CONCLUSION: Genistein is able to induce the apoptosis of transplanted tumor cells.This apoptosis may be mediated by down-regulating bcl-2 and up-regulating bax mRNA and its protein.     

Apoptosis of alveolar wall cells is involved in papain-induced rat pulmonary emphysema

AIM: To observe the apoptosis of alveolar wall cells in the papin-induced rat emphysema, and to explore the role of apoptosis of alveolar wall cells in the pathogenesis of emphysema. METHODS: Rats were randomly divided into three groups: normal control group, papain only group, papain+irradiation group. Morphology of lung tissues was assessed. TUNEL assay was used to determine the apoptotic cells. Immunohistochemistry was performed to determine the expression of PCNA, Bax and SP-C in the lung alveolar wall cells. SP-C immunofluorescence stainging was performed to identify the type II alveolar cells in the TUNEL positive cells. RESULTS: Destruction of alveolar wall and loss of the alveolar unit were observed in papain only group and papain+irradiation group. There was significant difference between rats of papain+irradiation group and papain only growp in the mean linear interval (MLI), the number of alveolar counted per unit area (MAN) and mean alveoli area (MAA), respectively (P<0.01). The proliferation index(PI), apoptosis index (AI) and the percentage of Bax in the papain only group and papain+irradiation group were significantly greater than that in the normal control group (P<0.01). However, the percentage of SP-C positive cells was significantly lower in the papain only group and papain+irradiation group compared with the normal control (P<0.01). Moreover, the PI, AI and the percentage of Bax in the papain+irradiation group was higher than those in the papain only group. The percentage of SP-C positive cells in the papain+irradiation group was lower than that in the papain only group. Most of the TUNEL-positive cells expressed the SP-C. CONCLUSION: Apoptosis of rat alveolar wall cells, especially type-II cell may be involved in the pathogenesis of emphysema. Upregulation of Bax expression may lead to alveolar wall cells apoptosis in papain-induced emphysema.     

Effects of andrographolide on invasion and apoptosis of ovarian cancer SKOV-3 cells

AIM:To investigate the effects of andrographolide on the invasion and apoptosis of ovarian cancer cell line SKOV-3,and to explore the possible mechanisms.METHODS:SKOV-3 cells were treated with different concentrations (0,5,10,20 or 40 μmol/L) of andrographolide for different time (12,24,36 or 48 h),and then the cell viability was determined by CCK-8 assay.The cell invasion ability was analyzed by Transwell assay and cell apoptosis was detected by TUNEL staining.The protein levels of p-PI3K,p-Akt and p-mTOR were examined by Western blot.RESULTS:The results of CCK-8 assay revealed that andrographolide inhibited the growth of SKOV-3 cells in a dose-and time-dependent manner.Treatment with andrographolide at 20 μmol/L for 36 h significantly decreased the invasion ability of SKOV-3 cells,while increased cell apoptosis.In addition,the protein levels of p-PI3K,p-Akt and p-mTOR were reduced after andrographolide treatment.CONCLUSION:Andrographolide inhibits the growth and invasion of ovarian cancer SKOV-3 cells by suppression of PI3K/Akt/mTOR signaling pathway.