Journal: Cancer Cell International

Article Title: Overexpression of trefoil factor 3 (TFF3) contributes to the malignant progression in cervical cancer cells

doi: 10.1186/s12935-016-0379-1

Figure Lengend Snippet: Forced expression of TFF3 in cervical cancer cells modulates the expression of malignant progression-related gene markers. a Determination of endogenous expression of TFF3 protein by Western blot in the cervical cancer cells lines CaSki, SiHa, Me180 and Hela and human non-tumor keratinocyte line HaCaT. b Semi-quantitative RT-PCR analysis was used to assess the mRNA levels of TFF3 in SiHa and Hela cells with either forced or depleted expression of TFF3 as described in “ ” section. c Western blot analysis was used to assess the protein levels of TFF3 in SiHa and Hela cells with either forced or depleted expression of TFF3. d , e Quantitative PCR analysis quantifying the change in expression of various genes associated with malignant progression in SiHa-TFF3/Hela cells. Change in gene expression is expressed as fold difference, respectively. Fold change values are representative of three independent experiments

Article Snippet: Human cervical cancer cell lines SiHa, CaSki, Hela, Me180 and human non-tumor keratinocyte line HaCaT were obtained from Nanjing KeyGen Biotech Co, Ltd (Nanjing,China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Oncology Reports

Article Title: Kindlin-2 suppresses cervical cancer cell migration through AKT/mTOR-mediated autophagy induction

doi: 10.3892/or.2020.7603

Figure Lengend Snippet: Kindlin-2 induces autophagy in cervical cancer cells. (A) Western blot results of kindlin-2 expression in cervical cancer cell lines. (B) Western blot results of kindlin-2 in the indicated cells. (C) Expression levels of p62, LC3, and GAPDH in the indicated cell lines. Cells were cultured in normal medium or EBSS-containing medium for 6 h. (D) Representative fluorescence images of CaSki and SiHa cells transfected with mRFP-GFP-LC3 lentivirus (magnification, ×630). (E) GFP and RFP puncta were counted in 30 cells. *P<0.05. NC, negative control; OE, overexpression; KD, knockdown; GFP, green fluorescent protein; RFP, red fluorescent protein; EBSS, Earle's balanced salts solution.

Article Snippet: The SiHa human cervical cancer cell line was purchased from Shanghai GeneChem Co., Ltd., and CaSki and C-33A human cervical cancer cells were purchased from Zhongqiaoxinzhou Biotech.

Techniques: Western Blot, Expressing, Cell Culture, Fluorescence, Transfection, Negative Control, Over Expression, Knockdown

Journal: Oncology Reports

Article Title: Kindlin-2 suppresses cervical cancer cell migration through AKT/mTOR-mediated autophagy induction

doi: 10.3892/or.2020.7603

Figure Lengend Snippet: Kindlin-2 induces autophagy by inactivating the AKT/mTOR pathway. (A) Protein levels of AKT, p-AKT, mTOR, p-mTOR, and GAPDH in the indicated CaSki and SiHa cells as detected by western blotting. (B) The indicated CaSki cells were treated with SC79 (20 µM) for 24 h and the protein levels of AKT, p-AKT, p62, LC3, and GAPDH were determined by western blotting. (C) The indicated SiHa cells were treated with MK2206 (5 µM) for 24 h and protein levels of AKT, p-AKT, p62, LC3 and GAPDH were detected by western blotting. *P<0.05. NC, negative control; OE, overexpression; KD, knockdown; ns, not significant.

Article Snippet: The SiHa human cervical cancer cell line was purchased from Shanghai GeneChem Co., Ltd., and CaSki and C-33A human cervical cancer cells were purchased from Zhongqiaoxinzhou Biotech.

Techniques: Western Blot, Negative Control, Over Expression, Knockdown

Journal: Oncology Reports

Article Title: Kindlin-2 suppresses cervical cancer cell migration through AKT/mTOR-mediated autophagy induction

doi: 10.3892/or.2020.7603

Figure Lengend Snippet: Kindlin-2 impairs the migration of cervical cancer cells by inducing autophagy. CaSki cells were transfected with kindlin-2 lentivirus or NC lentivirus and then treated with 5 mM 3-MA. SiHa cells were transfected with kindlin-2 RNAi or NC lentivirus and then treated with 20 nM RAPA. Wound healing and Transwell migration assays were performed to detect cell migration under different treatments. (A and B) Representative images (magnification, ×100) and quantification of CaSki cells subjected to the indicated treatments after scratch wounding. (C and D) Representative images (magnification, ×100) and quantification of SiHa cells subjected to the indicated treatments after scratch wounding. The wound area was evaluated using ImageJ software. (E) Representative images (magnification, ×200) and quantification of migrated (F) CaSki and (G) SiHa cells. Migrated cells were counted in five random fields. *P<0.05, **P<0.01, ***P<0.001. NC, negative control; OE, overexpression; KD, knockdown; 3-MA, 3-methyladenine; RAPA, rapamycin.

Article Snippet: The SiHa human cervical cancer cell line was purchased from Shanghai GeneChem Co., Ltd., and CaSki and C-33A human cervical cancer cells were purchased from Zhongqiaoxinzhou Biotech.

Techniques: Migration, Transfection, Software, Negative Control, Over Expression, Knockdown

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 is highly expressed in CC tissues and cells. (A) Heatmap of differentially expressed circRNAs in CC tissues and normal tissues according to the online data set (GSE102686). (B) Expression level of hsa_circ_0101119 was detected via reverse transcription-quantitative PCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and normal human cervical epithelial cell line, HcerEpic. **P<0.01 vs. HcerEpic cells. (C) Expression level of hsa_circ_0101119 in CC tissues and normal tissues, according to the online data set (GSE102686). circRNA/circ, circular RNA.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 recruits EIF4A3 to inhibit TCEAL6 expression in CC. (A) Bioinformatics was used to predict the interaction probabilities of the RNA-binding protein EIF4A3 with hsa_circ_0101119. Predictions with probabilities >0.5 were considered ‘positive’, suggesting that the corresponding RNA and protein are likely to interact. (B) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated hsa_circ_0101119 in SiHa and HeLa cell lysates. (C) Pull down assay indicated that biotin-labeled hsa_circ_0101119 interacted with EIF4A3. (D) Bioinformatics was used to predict the interaction probabilities of EIF4A3 with TCEAL6. (E) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated TCEAL6 in SiHa and HeLa cell lysates. (F) Expression levels of EIF4A3 and TCEAL6 were detected via RT-qPCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and a normal human cervical epithelial cell line, HcerEpic. (G) Expression levels of EIF4A3 and TCEAL6 in CC tissues and normal tissues, according to the analysis of TCGA. (H) Correlation between EIF4A3 and TCEAL6 in CC samples from TCGA. (I) After transfection with sh-EIF4A3, RT-qPCR was used to detect EIF4A3 expression in SiHa and HeLa cells. (J) After transfection with sh-EIF4A3, western blotting was performed to detect the expression level of TCEAL6 in SiHa and HeLa cells. (K) After co-transfection with si-hsa_circ_0101119 and sh-EIF4A3, western blotting was performed to measure the expression level of TCEAL6 in SiHa and HeLa cells. (L) A proposed model whereby hsa_circ_0101119 sequesters EIF4A3 away from TCEAL6 mRNA, in turn suppressing TCEAL6 mRNA translation. **P<0.01 vs. IgG group (B and E); *P<0.05, **P<0.01 vs. HcerEpic cells group (F); *P<0.05 vs. normal tissues group (G); **P<0.01, vs. sh-NC group (I and J); **P<0.01 vs. sh-NC group, ## P<0.01, vs. si-hsa_circ group. (K) RIP, RNA immunoprecipitation; RT-qPCR, reverse transcription-quantitative PCR; TCGA, The Cancer Genome Atlas; sh, short hairpin RNA; NC, negative control; si, small interfering RNA; circ, circular RNA; EIF4A3, eukaryotic initiation factor 4A-3; TCEAL6, transcription elongation factor A-like 6; T, tumor; N, normal; CC, cervical cancer.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, RNA Binding Assay, Pull Down Assay, Labeling, Quantitative RT-PCR, Transfection, Western Blot, Cotransfection, Immunoprecipitation, Real-time Polymerase Chain Reaction, shRNA, Negative Control, Small Interfering RNA