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@#[Abstract] Objective: To investigate the effect of cytokeratin 13 (CK13) on radio-sensitivity of human nasopharyngeal carcinoma HNE1 cell line and its mechanism. Methods: HNE1 cells were divided into control group, anti-CK13#a group (CK13 knockdown), anti-CK13#b group (CK13 knockdown), control+sirolimus group (100 nmol/L sirolimus treatment for 1 h), and anti-CK13#a + sirolimus group (100 nmol/L sirolimus treatment for 1 h). After irradiation treatment (200 cGy/min irradiation for 5 min), cell proliferation in each group was measured by CCK-8 assay. Cell apoptosis rate in each group was determined by Flow cytometry. Expression of PI3K/AKT/mTOR signaling pathway related PTEN gene was detected by qPCR, and WB was used to detect the expressions of PI3K/AKT/mTOR signaling pathway related proteins. Results: In the case of radiotherapy, as compared with the control group, the proliferation of HNE1 cells after CK13 knockdown was significantly enhanced (P<0.01) while the apoptosis rate was significantly reduced (P<0.01), the contents of caspase-3 and γH2AX as well as the protein lever of PTEN in cells were significantly decreased, while the expressions of p-AKT and p-S6K were significantly increased (all P<0.01). Interestingly, additional treatment with sirolimus (PI3K/AKT/mTOR signaling pathway inhibitor) could rescue the accelerated cell proliferation and decreased cell apoptosis caused by CK13 knockdown (all P<0.05). Conclusion: CK13 knockdown can enhance the activity of PI3K/AKT/mTOR signaling pathway by down-regulating PTEN, and ultimately reduce the radio-sensitivity of nasopharyngeal carcinoma HNE1 cells.
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Objective To investigate the effect of anti-human immunoglobulin M (IgM) on proliferation,apoptosis,cell cycle and tumor formation in human nasopharyngeal carcinoma HNE-1 cell line in vitro and in vivo.Methods After treatment with anti-human IgM antibody,proliferation of HNE-1 cells was observed by cell proliferation inhibition assay,apoptosis and cell cycle of HNE-1 cells were detected by flow cytometry,and apoptotic cells were detected by TUNEL staining.Nude mouse models were constructed,and were injected intraperitoneally with anti-human IgM antibodies (once every 3 days).The growth of transplanted tumor was observed once every 4 days.After the fifth injection,the expression levels of IgM and gp96 protein in transplanted tumor were observed by immunohistochemical method (streptavidin-peroxidase conjugated method,SP).Results MTS assay showed that anti-human IgM antibody can significantly inhibit the proliferation of HNE-1 cells in concentration-and time-dependent manner (P＜0.05).Flow cytometry showed that the anti-human IgM antibody promoted a significant decrease in percentage of cells in G1 phase,a significant increase in percentage of cells in S phase,and a significant increase in apoptotic rate of HNE-1 cells (P＜0.05).TUNEL staining showed that the anti-human IgM antibody promoted apoptosis of HNE-1 cells (P＜0.01).Transplantation tumor experiment showed that anti-human IgM antibody can significantly inhibit the volume and weight of transplanted tumor (P＜0.05).The immunohistochemistry showed that the expression levels of IgM and gp96 proteins in mouse transplanted tumors after intraperitoneal injection with anti-human IgM antibodies were significantly lower than those of the control group (P＜0.05).Conclusion The anti-human IgM anti-body could effectively inhibit the proliferation of HNE-1 cells,promote apoptosis,and arrest cell cycle.Anti-human IgM antibody could also inhibit the growth of transplanted tumor in nude mouse,which might be related to inhibition of the expressions of IgM and gp96 proteins.
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OBJECTIVE:To study the reversal mechanism of oxymatrine on multidrug resistance in the NPC HNE-1(200) cell line. METHODS: The multidrug resistant HNE-1(200) subline was derived from an HNE-1 cell line after exposure to fractionated X-irradiation. The expression of P-gp was tested by immunocytochemical method and western blot and that of mdr1 mRNA was measured by RT-PCR. RESULTS: After 24-hour treatment with oxymatrine, both the immunocytochemical method and western blot showed a decrease in the expression of P-gp. However, RT-PCR measurement showed that the expression of mdr1 mRNA of HNE-1 and HNE-1(200) was nonsignficant. CONCLUSION: Oxymatrine can reverse the multidrug resistance of NPC HNE-1(200) cell line to some degree by down-regulating the expression of P-gp. However, this medicine did no effect on the expression of mdr1 mRNA, suggesting the up-regulation of the expression of P-gp is under the control of multiple mechanism.
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Objective:To observe the effects of different concentrations of TRAIL and chemotherapeutic drugs(5-FU,DDP) on proliferation and apoptosis of HNE-1 cell lines ,both singlely and jointly.Methods:MTT was used to detect the inhibition rate.FCM was used to detect the apoptosis rate of cell.Results:The inhibition rate and apoptosis rate of combination use of TRAIL and 5-FU,DDP were significantly increased,compared with those of single use of TRAIL.Conclusion:The inhibition of proliferation and apoptosis of HNE-1 cell line can be induced by TRAIL,and the effects were higher by TRAIL combined with 5-FU and DDP.
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OBJECTIVE: To study the joint action of tea polyphenols and common anti-tumor drugs(vincristine,pingyangmycin,5-Fu,cisplatin) in inhibiting cell proliferation of the multiresistant HNE-1(200) cell line in patients with nasopharyngeal carcinoma after radiographic exposure.METHODS: The low toxic dosage of tea polyphenols and 4 kinds of anti-tumor drugs were optimized by MTT method.Then the inhibition ratio of multiresistant HNE-1(200) cell line treated by tea polyphenols and anti-tumor drugs waere detected.RESULTS: The low toxic dosage of tea polyphenols was 0.50 mg?mL-1.When the anti-tumor drugs were used in combination with tea polyphenols(0.50 mg?mL-1) as compared without,the inhibition ratio on HNE-1(200) cell line was increased by 100%～300%.CONCLUSIONS: Addition of tea polyphenols to anti-tumor drugs showed remarkable inhibitory effect on cell proliferation of HNE-1(200) cell line in patients with nasopharyngeal carcinoma,suggesting the reverse effect of tea polyphenols on resistant HNE-1(200) cell line.
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Objective:To investigate the effects of oxymartine on cell growth and the changes of P-glycoprotein in HNE-1 cell line after exposed to fractionated X-irradiation.Methods:By MTT,the cell proliferation was observed both before X-irradiation and after X-irradiation conditions with the extent of vincristine,oxymartine and verapamil respectively.P-glycoprotein expression was tested by immunohistochemistory method SP.Results:(1)Vincristine,oxymartine and verapamil were capable of inhibiting cell growth on HNE-1 cell line and fractionated X-irradiated HNE-1(200)cell subline;(2)Vincristine was less inhibitory on cell growth in HNE-1(200)cell subline than HNE-1 cell line,and the difference had statistic significance;(3)P-glycoprotein expression in HNE-1 cell line was negative while HNE-1(200)cell subline positive;oxymartine with low inhibitory concentration could increase the expression of P-glycoprotein in HNE-1(200) cell subline,which is similar to that of verapamil.Conclusion:(1)Oxymartine is capable of inhibiting cell growth on HNE-1 cell line and fractionated X-irradiated HNE-1(200)cell subline;(2)HNE-I(200)cell subline is resistant to vincristine;(3)Oxymartine with low inhibitory concentration could increase the expression of P-glycoprotein in HNE-1(200)cell subline.which is similar to that of verapamil.