|
|
|
| Effect of cyclooxygenase inhibitor NS-398 combined with Oxaliplatin on growth inhibition and apoptosis induction of HeLa cells in cervical cancer |
| WANG Jing1 JIN Yan2 DONG Shanglin3 WU Xin4 |
1.Department of Gynaecology and Obstetrics, the First Affiliated Hospital of Hebei North University, Heibei Province, Zhangjiakou 075000, China;
2.Life Science Research Center, Hebei North University, Heibei Province, Zhangjiakou 075000, China;
3.the First Affiliated Hospital of Hebei North University, Heibei Province, Zhangjiakou 075000, China;
4.Department of Pathology, Hebei North University, Heibei Province, Zhangjiakou 075000, China |
|
|
|
Abstract Objective To explore the effect of cyclooxygenase inhibitor NS-398 combined with Oxaliplatin (L-OHP) on growth inhibition and apoptosis induction of HeLa cells in cervical cancer. Methods HeLa cells were cultured in vitro. CCK-8 method was used to detect the inhibitory effect of different concentrations of NS-398 (0, 30, 60 μmol/L) and L-OHP (0.0, 2.5, 5.0, 10.0 mg/L) on HeLa cells alone or in combination. HeLa cells were divided into: control group, NS-398 group (60 μmol/L), L-OHP group (10 mg/L) and combined group (NS-398 60 μmol/L+L-OHP 10 mg/L). The morphological changes of HeLa cells were observed by inverted microscope. The apoptosis of HeLa cells was observed by Rhodamine staining, and the early apoptosis rate of HeLa cells was detected by flow cytometry. The ladder strip was detected by DNA agarose gel electrophoresis. Results CCK-8 analysis showed that compared with untreated group, the cell growth inhibition rate in NS-398 single drug group and L-OHP single drug group were higher; growth inhibition rate in combined group was higher than that in NS-398 single drug group and L-OHP single drug group (all P ＜ 0.01). The cells in control group were in good condition, with fusiform shape, adherent growth, clear cell edges and intact cell membrane. In NS-398 group, the volume of cells shrank and became round, granular substances appeared in the cytoplasm, and a small number of cells fell off from the bottom of the bottle and floated in the culture medium. The cells of L-OHP group lost their normal morphology and shrank, and dead cells could be seen in the culture medium. A large number of dead cells and cell debris could be seen in the culture medium of the combined group, with only a few cells sticking to the bottom of the bottle, with different degrees of shrinkage, partial cell rupture, and cell contents overflow. Rodamine 123 staining result showed that, the control group had no fluorescence. The fluorescence intensity of NS-398 group, L-OHP group and combination group was enhanced, especially the combination group. The results of flow cytometry showed that the apoptosis rate of NS-398 group and L-OHP group were significantly higher than those of control group; the apoptosis rate of combined group was higher than that of NS-398 group and L-OHP group (all P ＜ 0.01). DNA electrophoresis showed that there were obvious ladder-like bands in combined group, while there were obvious dispersion phenomena in NS-398 group and L-OHP group. In control group, neither dispersion nor ladder band appeared. Conclution NS-398 can enhance the inhibitory effect of L-OHP on the growth of HeLa cells and induce apoptosis.
|
|
|
|
|
|
[1] Shrestha AD，Neupane D，Vedsted P，et al. Cervical cancer prevalence，incidence and mortality in low and middle income countries：a systematic review [J]. Asian Pac J Cancer Prev，2018，19（2）：319-324.
[2] Hoppenot C，Stampler K，Dunton C. Cevrical cancer screening in high-and low-resource countries：Implications and new developments [J]. Obstet Gynecol Surv，2012，67（10）：658-667.
[3] Dai F，Chen G，Wang Y，et al. Identification of candidate biomarkers correlated with the diagnosis and prognosis of cervical cancer via integrated bioinformatics analysis [J]. Onco Targets Ther，2019，12：4517-4532.
[4] Chen W，Zhang R，Baade PD，et al. Cancer statistics in China，2015 [J]. CA Cancer J Clin，2016，66（2）：115-132.
[5] Lai CH. Management of recurrent cervical cancer [J]. Chang Gung Med J，2004，27（27）：711-717.
[6] Mauri G，Gori V，Bonazzina E，et al. Oxaliplatin retreatment in metastatic colorectal cancer：Systematic review and future research opportunities [J]. Cancer Treat Rev，2020， 91：102112.
[7] Christopher NP，Manuel H，Ignacio GL，et al. From state-of-the-art treatments to novel therapies for advanced-stage pancreatic cancer [J]. Nat Rev Clin Oncol， 2020，17（2）：108-123.
[8] Salvatore G，Francesco E，Francesca P，et al. Gercumin synergizes the action of 5-fluorouracil and oxaliplatin against chemoresistant human cancer colon cells [J]. Biochem Biophys Res，2020，522（1）：95-99.
[9] Shepard G, Arrowsmith ER, Murphy P，et al． A phase Ⅱ study with lead-in safety cohort of 5-fluorouracil，oxaliplatin，and lapatinib in combination with radiation therapy as neoadjuvant treatment for patients with localized HER2-positive esophagogastric adenocarcinomas [J]. Oncologist，2017，22（10）：1152-1198.
[10] 马振华，姜海涛，张燕，等.NS-398联合光动力疗法抑制环氧合酶-2、血管内皮生长因子-C对胆管癌QBC939细胞凋亡的影响[J].中华实验外科杂志，2020， 37（11）：2062-2066.
[11] Palumbo P，Lombardi F，Augello FR，et al. Biological effects of selective COX-2 inhibitor NS398 on human glioblastoma cell lines [J]. Cancer Cell Int，2020，20：167.
[12] Ferlay J，Shin HR，Bray F，et al. Estimates of worldwide burden of cancer in 2008：GLOBOCAN 2008 [J]. Int J Cancer，2010，127（12）：2893-2917.
[13] Chen MH，May BH，Zhou IW，et al. Integrative medicine for relief of nausea and vomiting in the treatment of colorectal cancer using oxaliplatin-based chemotherapy：a systematic review and meta-analysis [J]. Phytother Res，2016，30（5）：741-753.
[14] 张强，李邵青，刘薇，等.基于糖基化修饰的铂类抗肿瘤化合物研究进展[J].中国药学杂志，2020，55（17）：1409-1414.
[15] Pasetto LM，D’Andre MR，Rossi E，et al. OxaliPlatin-related neurotoxieity：how and why？ [J]. Crit Rev OncolHematol，2006，59（2）：159-168.
[16] Raymond E，Faivre S，Chaney S，et al. Cellular and molecular Pharmacology of Oxaliplatin [J]. Mol Cancer Ther，2002，1（3）：227-235.
[17] Micheli L，Mattoli L，MaidecchiA，et al. Effect of Vitis vinifera hydroalcoholic extract against oxaliplatin neurotoxicity：in vitro and in vivo evidence [J]. Sci Rep，2018， 8（1）：14364.
[18] 武玉，王益民，赵敏，等.MiR-885对结直肠癌细胞奥沙利铂化疗敏感性的影响[J].中国肿瘤，2020，29（10）：799-804.
[19] Soko?覥owski G，Ba?覥dys-Waligórska A，Trofimiuk M，et al. Expression of cyclooxygenase-2（COX-2）in pituitary tumours [J]. Med Sci Monit，2012，18（4）：CR252-CR259.
[20] Soares CD，Hernandez-Guerrero JC，Andrade BA，et al. Comparative expression of cyclooxygenase 2 and Ki67 in amelanotic and conventional oral melanomas [J]. Med Oral Patol Oral Cir Bucal，2020，25（6）：e728-e731.
[21] Kim HJ，Wu HG，Park IA，et al. High cyclooxygenase-2 expression is related with distant metastasis in cervical cancer treated with radiotherapy [J]. Int J RadiatIncolBiol Phys，2003，55（1）：16-20.
[22] Chen YJ，Wang LS，Wang PH，et al. High cyclooxygenase-2 expression in cervical adenocarcinomas [J]. Gynecol Oncol，2003，88（3）：379-385.
[23] Ye Y，Wang X，Jeschke U，et al. COX-2-PGE2-EPs in gynecological cancers [J]. Arch Gynecol Obstet，2020， 301（6）：1365-1375.
[24] Wu QM，Li SB，Tong Q，et al. Inhibitory effects of a cyclooxygenase-2 inhibitor，NS398，on cancer cells [J]. Chin J Pathophysiol，2006，22（11）：2231-2234.
[25] Fornetti J，Jindal S，Middleton KA，et al. Physiological COX-2 expression in breast epithelium associates with COX-2 levels in ductal carcinoma in situ and invasive breast cancer in young women [J]. Am J Pathol，2014， 184（4）：1219-1229. |
|
|
|
