Silencing NOB1 enhances doxorubicin antitumor activity of the papillary thyroid carcinoma in vitro and in vivo
Authors: Shuai Xue
Guang Chen
Xiao-Νan Zhang
Bing-Fei Dong
Jia Liu
Zhe Han
Pei-Song Wang
Pei-You Ren
Journal: Oncology Reports (2015)
Discipline: CLINICAL MEDICINE
Keywords: Doxorubicin; NOB1; Papillary thyroid carcinoma; Resistance; Adenovirus
Affiliations: Department of Thyroid Surgery, The First Bethune Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.

Abstract:  Doxorubicin (DOX), a broad‑spectrum anthra-cyclin, is in wide clinical use for the treatment and prevention of thyroid cancer. However, the effectiveness of the treatment remains limited due to inherent tumor resistance to DOX. Results of a previous study demonstrated that downregulation of NIN1/RPN12 binding protein 1 homolog (NOB1) expression via adenovirus expression vector carrying NOB1 siRNA (Ad/sh-NOB1) induced cancer apoptosis and increased the radiosensitivity of papillary thyroid carcinoma (PTC) cells. However, whether knockout NOB1 can decrease DOX resistance remains unclear. Therefore, in the present study, the effect of Ad/sh-NOB1 infection, independently or in combination with DOX, was determined in a PTC cell line to identify more effective therapeutics against PTC cancer. Furthermore, tumor growth ability in nude mice was determined to identify the combination treatment effect in tumorigenesis in vivo. The results showed that Ad/sh-NOB1 combined with DOX treatment in PTC cells significantly suppressed proliferation, colony formation, migration and invasion, and induced cell apoptosis and arrest in the G0/G1 stage as compared to Ad/sh-NOB1 or DOX monotherapy. We also found that this combination suppressed the tumor growth of a nude mouse model as compared to Ad/sh-NOB1 or DOX monotherapy. In addition, Ad/sh-NOB1 combined with DOX treatment significantly increased activation of the p38 MAPK pathway, which may contribute to inhibition of PTC cell growth and decreased DOX resistance. Taken together, the experimental results indicate that Ad/sh-NOB1 combined with DOX treatment is a potential drug candidate for the treatment of papillary thyroid carcinoma. 

Retracted: Yes

PubPeer comments:
 *[left]* Fig 5.  Effect of DOX and Ad sh-NOB1 alone or in combination on the p38MAPK signaling pathway in TPC-1 cells. Phosphorylation activation pattern of p38MAPK, ERK1 2 and JNK was measured by western blotting 12 h after treatment with DOX and Ad sh-NOB1 alone or in combination in TPC-1 cells. GAPDH was used as an internal control. *[right]* Fig 4d from [Li et al (2016)] .  AKT3 and p-AKT protein expression in K1 cells transfected with miR-29a mimic or miR-Ctrl were determined by Western blot. qRT-PCR and Western blot data were normalized to GAPDH . ![file] One band seems to be the same (after a 180 deg flip). No authors in common. , 
 *[left]* Fig 5 again. *[right]* Fig 4 from [Lin et al (2017)] .  miR-215 activate MAPK signal pathway. The p38MAPK, phosphorylated-p38MAPK (p-p38MAPK), ERK, p-ERK, JNK and p-JNK protein expression were measured in SKOV3 cells transfected with miR-215 and miR-NC or infected with Lv sh-NOB1 or Lv shScramble by Western blot . I have concerns. ![file] UPDATE: A few more bands coloured now, to show affiliations to Fig 4D of [Li et al (2016)] .  IGF1R, AKT, p-AKT, ERK, and p-ERK protein in SKOV3 cells transfected with miR-494 mimic were determined by western blot . ![file] , 
 Fig 2C.  Western blot analysis of cyclin D1 and D3, and p21 protein expression following treatment with DOX and Ad sh-NOB1 alone or in combination. GAPDH was used as an internal control. ![file] The loading control was beta-Actin in *[left]* Fig 3E of [Yang et al (2015)] ,  western blot analysis of MMP-2 and MMP-9 protein expression following treatment with SOR and CDDP alone or combination . ![file] See also Fig 3B of [Sang et al (2016)](http: www.ijcem.com files ijcem0019312.pdf).  PIK3R3 on ... protein level were determined in 786-O cells transfected with si-PIK3R3 or si-NC by... western blot ; and other sightings in those threads. ![file] , 
 *[right]* Fig 6A.  Graphs of the tumor tissue from the different groups . *[left]* Fig 6A from [Qu et al (2015)] .  Images of tumor tissue with different plasmid treatments collected after sacrifice at day 21. ![file] For comparison, *[left]* Fig 7A from [Zhang et al (2015)] .  Images of tumor tissue with different groups collected after sacrifice at day 28 . *[right]* Fig 7A from [Tian et al (2015)] .  Images of tumor tissues in the different mouse groups collected after sacrifice at day 28. ![file] I have concerns. ----------------------------------------- UPDATE: Fig 5A from [Yang et al (2015)] .  Images of tumor tissue from different treatments collected after sacrifice at day 22. ![file] I have not tried to mark up the CDDP+SOR or the SOR bands, for fear of obscuring the change in texture around the 2nd tumor in the former, or the 5th tumor in the latter. , 
 *[left]* Fig 2A from [Liu et al (2015)] .  Effect of DOX and Ad sh-NOB1 alone or in combination on cell cycle of TPC-1 cells. (A) Cell DNA content distribution in each phase . *[upper right]* Fig 2C from [Song et al (2015)] .  Determination of HeLa cell cycle distribution by PI staining after transfection with miR-133a or miR-Ctrl . *[lower right]* Fig 2d from [Zhang et al (2015)] .  The cell cycle was detected by PI staining after being transfected with miR-133a or miR-NC. ![file] The same panels keep appearing. , 
 Clockwise from top left: - Fig 3A from [Liu et al (2015)] .  Effects of miR-222 inhibitor on HepG2 cell cycle distribution. (A) Cell DNA content distribution in each phase. - Fig 3A from [Zhang et al (2015)] .  Effects of microRNA-145 (miR-145) on multiple myeloma (MM) cell cycle distribution. (A) Cell DNA content distribution in each phase. - Fig 3A from [Li et al (2015)] .  Enforced expression of miR-338-3p induced cell cycle distribution and cell apoptosis in EC9706 cells. (A) Cell DNA content distribution in each phase. - Fig 3A from [Zhang et al (2015b)] ,  miR-137 induces changes in cell cycle distribution and cell apoptosis in SKOV3 cells. (A) Cell DNA content distribution in each phase ![file] The same panels keep appearing. , 
 Whoops, a box in Comment #5 is the wrong color. Trying again: *[left]* Fig 2A from [**Liu et al (2015)**] .  Effect of DOX and Ad sh-NOB1 alone or in combination on cell cycle of TPC-1 cells. (A) Cell DNA content distribution in each phase . *[upper right]* Fig 2C from [Song et al (2015)] .  Determination of HeLa cell cycle distribution by PI staining after transfection with miR-133a or miR-Ctrl . *[lower right]* Fig 2d from [Zhang et al (2015)] .  The cell cycle was detected by PI staining after being transfected with miR-133a or miR-NC. ![file] The same panels keep appearing. --------------------------------------------------- Clockwise from upper left: - Fig 2D from [Pang et al (2015)] .  cell cycle progression ... determined in HepG2 cells transfected with the miR-154 mimic or miR-NC. - Fig 2D from [Zhao et al (2017)] .  cell cycle stage ... assessed in MM1S cells transfected with miR-144-3p mimic or miR-NC. - Fig 2D from [Li et al (2016)] .  The cell cycle procession was detected by flow cytometry after transfection with miR-494 or miR-NC . - Fig 2C from [Luan et al (2015)] .  Cell cycle was measured in glioma cells at posttransfection by flow cytometry . ![file] The same panels keep appearing. --------------------------------- Fig 2D from [Zhou et al (2016)] .  Cell cycle distribution of U2OS cells transfected with the miR-154 mimic was determined by flow cytometry . ![file] , 
 Retracted [1 March 2022] .  Following the publication of this paper, it was drawn to the Editorsu2019 attention by a concerned reader that certain of the western blotting assay data shown in Figs. 2 and 5, and the tumour images shown in Fig. 6A, were strikingly similar to data appearing in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had already been published elsewhere, or were already under consideration for publication, prior to its submission to *Oncology Reports*, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they agreed with the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. ,