MicroRNA-1179 regulates proliferation and chemosensitivity of human ovarian cancer cells by targeting the PTEN-mediated PI3K/AKT signaling pathway
Authors: Chen Rubin
Wu Yanting
Lu Liping
Guo Hui
Zhang Zhihong
Mao Anpeng
Shan Zhenxiu
Journal: Archives of Medical Science (2020)
Discipline: CLINICAL MEDICINE
Keywords: apoptosis; miR-1179; microRNA; ovarian cancer.
Affiliations: Department of Oncology, Gong'an County People's Hospital, Gong'an County, Hubei Province, China., Department of Radiology, Gong'an County People's Hospital, Gong'an County, Hubei Province, China.

Abstract:  Introduction: Owing to widespread roles of miRs, the dysregulation of their expression in human tissues has been linked with the development of several diseases such as cancer. The study was designed to investigate the role and therapeutic potential of miR-1179 in ovarian cancer. Material and methods: Proliferation rate was monitored by MTT assay. Transfections were performed using Lipofectamine 2000 reagent. Cell cycle apoptosis was detected by AO/EB and annexin V/PI staining. Expressions analysis was carried out by qRT-PCR and western blotting. In vivo evaluation was carried out in xenografted mouse models. Results: The results revealed that miR-1179 is considerably upregulated in ovarian cancer cell lines. Inhibition of miR-1179 triggers decrease in the viability via initiation of apoptotic cell death of ovarian PA-1 cancer cells. TargetScan analysis showed PTEN to be the main target of miR-1179 in PA-1 cells. Exploration of PTEN expression in ovarian cancer cell lines revealed up to 9-fold downregulation of PTEN. However, inhibition of miR-1179 in PA-1 cells resulted in upregulation of PTEN expression. In addition, overexpression of PTEN caused a reduction of PA-1 cell viability via induction of apoptotic cell death. However, silencing of miR-1179 could rescue the effects of miR-1179 inhibition on the proliferation of miR-1179. The miR-1179 suppression was accompanied by a significant decline in phosphorylation of PI3K and AKT expression in the PA-1 cells. The in vivo study showed that miR-1179 suppression inhibits the xenografted tumor growth. Conclusions: The results of this study indicate that miR-1179 may prove to be an important therapeutic target for ovarian cancer. 

Retracted: No

PubPeer comments:
 Fig 1E.  Suppression of miR-1179 inhibits proliferation of ovarian cancer cell lines:... annexin V PI staining of NC or miR-1179 inhibitor transfected PA-1 and SK-OV-3 Fig 3H.  miR-1179 exerts its effects by targeting PTEN... Annexin V PI staining of NC or pcDNA-PTEN transfected PA-1 ovarian cancer cells ![file] For comparison: *[left]* Figure 6 from [Liu et al (2019)] .  Annexin V PI assay showing percentage of apoptotic MDA-MB-231 cells at indicated concentrations of withaferin-A. *[right]* Fig. 2b from [Li et al (2020)] .  Annexin V PI staining showing the percentage of apoptotic U87 and U118 cells . ![file] Figure 3B from [Xu et al (2019)] .  miR-181 induces apoptosis in glioma cells. ...  (B) annexin V PI staining of miR-NC and miR-181 mimics transfected U87 and U118 cells showing induction of apoptosis. ![file] , 
 *[right]* Fig 1E from [Zhang et al (2020)] .  Suppression of miR-1179 inhibits proliferation of ovarian cancer cell lines ... DAPI, AO EB or annexin V PI staining of NC or miR-1179 inhibitor transfected PA-1 . *[left]* Figure 3 from [Yang, Cai Zho (2020)] .  A: DAPI staining, B: AO EB staining, and C: Annexin-V PI staining of miR-NC and miR-1 mimics transfected MBA-MD-231 cells. The Figure depicts that miR-1 overexpression induces apoptosis in human breast MBA-MD-231 cells. ![file] , 
 *[above]* Fig 2A from [Zhang et al (2020)] .  AO EB staining showing the induction of apoptosis at in U87 and U118 cells . *[below]* Figure 2C from [Li et al (2019)] .  miR-23 overexpression inhibits the proliferation of gastric cancer cells. ... C: AO EB staining showing apoptosis of miR-NC and miR-23 mimics transfected AGS cells . ![file] Overlap marked. , 
 #2 I forgot to compare *[left]* the other half of Fig 1E, for SK-OV-3 cells, with *[right]* Figure 3A,B from [Xu et al (2019)] .  miR-181 induces apoptosis in glioma cells. (A) DAPI staining and (B) annexin V PI staining of miR-NC and miR-181 mimics transfected U87 and U118 cells showing induction of apoptosis . ![file] , 
 *Material and methods* *Cell lines and culture conditions* *The human ovarian cancer cell lines (SK-OV-3, OVACAR-3, PA-1, SW-626, Caov3) and normal ovarian cell line (**SV40**) were obtained from Sun Yat-sen University Cancer Center (Guangzhou, China). Culturing of the cells was performed in RPMI 1640 medium (Invitrogen). The media was supplemented with 10% FBS and the incubation of the cells was carried out at 37u00b0C in a humidified atmosphere with 5% CO2.*  Normal ovarian cell line (SV40) is presumably one that has been immortalized with SV40. This seems unspecific. Surely the source would have stated the nature of the cells. If these cells are a standard product from Sun Yat-sen University Cancer Center, is there some kind of identifying code? ------------------------------------------ *Results* *miR-1179 is upregulated in ovarian cancer cell lines* *Gene expression analysis revealed miR-1179 to be significantly upregulated in the ovarian cancer tissues as compared to the adjacent normal tissues. The upregulation of miR-1179 in ovarian cancer tissues was observed to be up to more than 7-fold (Figure 1 A).* It seems that samples of ovarian cancer and normal tissue were obtained from 15 patients, but this is not elsewhere mentioned in the paper. The Methods section only describes the cultured cell-lines. The authors urgently need to provide basic demographics for the 15 tissue donors, where they were recruited, and ethical assurances that the donations were consenting and fully-informed. ,