"VSports" Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil VSports app下载. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

. 2015 Oct 15;5(11):3350-62.
eCollection 2015.

V体育官网 - MLN4924, a Novel NEDD8-activating enzyme inhibitor, exhibits antitumor activity and enhances cisplatin-induced cytotoxicity in human cervical carcinoma: in vitro and in vivo study

Wei-Chou Lin  1 , Kuan-Lin Kuo  2 , Chung-Sheng Shi  3 , June-Tai Wu  4 , Ju-Ton Hsieh  5 , Hong-Chiang Chang  5 , Shih-Ming Liao  5 , Chien-Tso Chou  5 , Chih-Kang Chiang  6 , Wei-Shuo Chiu  7 , Tzu-Yuan Chiu  4 , Yeong-Shiau Pu  5 , I-Lin Ho  5 , Zuo-He Wang  5 , Shih-Chen Chang  5 , Shing-Hwa Liu  6 , Yung-Ming Jeng  1 , Kuo-How Huang  5
Affiliations

MLN4924, a Novel NEDD8-activating enzyme inhibitor, exhibits antitumor activity and enhances cisplatin-induced cytotoxicity in human cervical carcinoma: in vitro and in vivo study

VSports app下载 - Wei-Chou Lin et al. Am J Cancer Res. .

"VSports" Abstract

MLN4924, an inhibitor of NEDD8 activating enzyme (NAE), has been reported to have activity against various malignancies. Here, we investigated the antitumor properties of MLN4924 and MLN4924 in combination with cisplatin on human cervical carcinoma (CC) in vitro and in vivo. Two human CC cell lines, ME-180 and HeLa, were used in this study. The cytotoxic effects of MLN4924 and/or cisplatin were measured by cell viability (MTT), proliferation (BrdU incorporation), apoptosis (flow cytometry with annexin V-FITC labeling), and the expression of cell apoptosis-related proteins (Western blotting). In vivo efficacy was determined in Nu/Nu nude mice with ME-180 and HeLa xenografts. The results showed that MLN4924 elicited viability inhibition, anti-proliferation and apoptosis in human CC cells, accompanied by activations of apoptosis-related molecules and Bid, Bcl-2 phosphorylation interruption, and interference with cell cycle regulators VSports手机版. Moreover, MLN4924 caused an endoplasmic reticulum stress response (caspase-4, ATF-4 and CHOP activations) and expression of other cellular stress molecules (JNK and c-Jun activations). Additionally, MLN4924 suppressed growth of CC xenografts in nude mice. Furthermore, we demonstrated that MLN4924 potentiated cisplatin-induced cytotoxicity in CC cells with activation of caspases. Consistently with this, MLN4924 significantly enhanced cisplatin-induced growth inhibition of CC xenografts. Together, these findings suggest that MLN4924 alone or in combination with cisplatin is of value in treating human CCs. .

Keywords: MLN4924; apoptosis; cervical carcinoma; neddylation. V体育安卓版.

PubMed Disclaimer

V体育2025版 - Figures

Figure 1
Figure 1
MLN4924 elicited inhibition of cell viability and apoptosis in human CC cells. A. Cullin-1 neddylation was completely inhibited by MLN4924. ME-180 and HeLa cells were exposed to different concentrations of MLN4924 or DMSO (as non-treated control) for 48 hours and then harvested to detect the expression of cullin-1 with a Western blot (GAPDH as control). B. The cells were treated with various concentrations of MLN4924 for 24 and 48 hours. Cell viability was assessed with an MTT assay. C. The cells were exposed to MLN4924 (500 nM) or DMSO (as the non-treated control) for 48 hours. Apoptotic cells were analyzed by FACS flow cytometry with annexin V-FITC/PI staining. The annexin V-positive cells (early apoptotic cells) are distributed in the lower-right panel; the late apoptotic cells, whose membranes are permeable to PI and annexin V staining, are distributed on the upper-right panel. D. Quantitative analyses of total apoptosis population following MLN4924 (500 nM) or DMSO treatments were performed. Data are presented as the means ± SD of at least three independent experiments. *P < 0.05 compared with the controls.
Figure 2
Figure 2
MLN4924 activated caspases, PARP, phospho-Histone H2A.X, and Bid and decreased Bcl-2 phosphorylation in human CC cells. Total cell lysates were harvested and analyzed by Western blotting with specific antibodies against (A) apoptosis-related molecules (PARP, cleaved caspase-3, -7 and -8), DNA damage response regulator (phospho-Histone H2A.X), (B) Bid, and phospho-Bcl-2 (Ser70). Results shown represent at least three independent experiments.
Figure 3
Figure 3
MLN4924 activated cellular stress and endoplasmic reticulum stress-related signaling in human CC cells. Cells were treated with several concentrations of MLN4924 or DMSO (as the non-treated control) for 48 hours. Cell lysates were harvested, and the expression levels of cell stress-related proteins (phospho-SAPK/JNK and phospho-c-Jun) and ER stress-related molecules (ATF-4, CHOP and caspase-4) were assessed via a Western blot. Results shown are representative of at least three independent experiments.
Figure 4
Figure 4
MLN4924 reduced cell proliferation and interfered cell cycle mediators in ME-180 and HeLa cell lines. A. Cells were exposed to MLN4924 (500 nM) or DMSO (as the non-treated control) for 48 hours. BrdU incorporation assay was adopted to determine the proliferation of human CC cells, and the cell proliferation of these two cell lines was decreased after MLN4924 treatments. B. After exposure to various concentrations of MLN4924 for 48 hours, the expression levels of cell cycle mediators, such as p21, p27, phospho-p53 (Ser15), phospho-Chk1 (Ser345) and phospho-Chk2 (Thr68), in ME-180 and HeLa cells were analyzed by Western blotting. Results represent at least three independent experiments.
Figure 5
Figure 5
MLN4924 significantly abolished the growth of CC xenografts in vivo. Nu/Nu nude mice bearing ME-180 and HeLa CC tumor xenografts were injected intraperitoneally with DMSO (as the non-treated control; ME-180: n = 7, HeLa: n = 8) or MLN4924 (10 mg/kg; ME-180: n = 8, HeLa: n = 7) once per day for 32 days. A. The tumor images represent excised tumors from each group. B. The tumor weights were compared between the MLN4924-treated and control (DMSO) groups on the last day of treatment. Tumor weights are presented as the mean ± S.D.; *P < 0.05. C. The tumor volumes measured during the treatment are presented as the responses. Tumor volumes are presented as the mean ± SEM; *P < 0.05.
Figure 6
Figure 6
MLN4924 potentiated cisplatin-induced cytotoxicity in human CC cells. A. ME-180 and HeLa cells were incubated with MLN4924 (250 nM) and different concentrations of cisplatin alone or in combination for 48 hours. Cell viability was assessed with an MTT assay. Quantitative analyses of cell viability are presented as the means ± SD of five independents experiments. *P < 0.05 compared with cisplatin treatment alone. B. The total cell lysates were harvested and analyzed by Western blotting with specific antibodies against cleaved caspase-3 and caspase-8 after treatment with DMSO (as the non-treated control), cisplatin, MLN4924, and cisplatin/MLN4924 combination for 48 hours. Combination treatment with cisplatin/MLN4924 resulted in higher expression levels of apoptosis-associated proteins (cleaved caspase-3 and caspase-8) than the cisplatin or MLN4924 treatment alone. Results represent at least three independent experiments.
Figure 7
Figure 7
The anti-tumor efficacy of the combination of cisplatin and MLN4924 in a xenograft model of human cervical cancer. Nu/Nu nude mice bearing ME-180 or HeLa xenograft tumors were treated with DMSO (as non-treated control), cisplatin (1.2 mg/kg), MLN4924 (6 mg/kg) or the cisplatin/MLN4924 combination (1.2 mg/kg cisplatin plus 6 mg/kg MLN4924) by intraperitoneal injection once per day for 32 days. A. The tumor images represent excised tumors from each group. B. The tumor weights of each group (DMSO (as non-treated control), cisplatin, MLN4924 and combined treatment) measured on the last day of treatment were compared. C. The tumor volumes assessed during the different treatments are presented as the response. Quantitative analyses of tumor volumes are presented as the means ± SEM (n = 8 for each group of ME-180; n = 7 for each group of HeLa) and *P < 0.05 compared with cisplatin treatment alone or MLN4924 treatment alone.
See this image and copyright information in PMC

References

    1. Arbyn M, Castellsagué X, de Sanjosé S, Bruni L, Saraiya M, Bray F, Ferlay J. Worldwide burden of cervical cancer in 2008. Ann Oncol. 2011;22:2675–2686. - PubMed
    1. Blank JL, Liu XJ, Cosmopoulos K, Bouck DC, Garcia K, Bernard H, Tayber O, Hather G, Liu R, Narayanan U, Milhollen MA, Lightcap ES. Novel DNA damage checkpoints mediating cell death induced by the NEDD8-activating enzyme inhibitor MLN4924. Cancer Res. 2013;73:225–234. - "V体育2025版" PubMed
    1. Bosch FX, Lorincz A, Muñoz N, Meijer CJ, Shah KV. The causal relation between human papillomavirus and cervical cancer. J Clin Pathol. 2002;55:244–265. - V体育平台登录 - PMC - PubMed
    1. Bossy-Wetzel E, Bakiri L, Yaniv M. Induction of apoptosis by the transcription factor c-Jun. EMBO J. 1997;16:1695–1709. - PMC - PubMed
    1. Brownell JE, Sintchak MD, Gavin JM, Liao H, Bruzzese FJ, Bump NJ, Soucy TA, Milhollen MA, Yang X, Burkhardt AL, Ma J, Loke HK, Lingaraj T, Wu D, Hamman KB, Spelman JJ, Cullis CA, Langston SP, Vyskocil S, Sells TB, Mallender WD, Visiers I, Li P, Claiborne CF, Rolfe M, Bolen JB, Dick LR. Substrate-assisted inhibition of ubiquitin-like protein-activating enzymes: the NEDD8 E1 inhibitor MLN4924 forms a NEDD8-AMP mimetic in situ. Mol Cell. 2010;37:102–111. - PubMed