cervical cancer cell lines c33a hpv negative  (ATCC)

Journal: Discover. Oncology

Article Title: miR-17 ~ 92 suppresses proliferation and invasion of cervical cancer cells by inhibiting cell cycle regulator Cdt2

doi: 10.1007/s12672-023-00775-3

Figure Lengend Snippet: miR-17 ~ 92 over expression decreases Cdt2 expression in cervical cancer cells. A RT-qPCR analysis of mRNA level of Cdt2, 48 h after transfection of miR-17 ~ 92 in HEK293T, C33A and SiHa cell lines B Western blot for analysis of Cdt2 protein expression level, 48 h after transfection of miR-17 ~ 92 in HEK293T, SiHa, HeLa and C33A cell lines. C Quantification of Cdt2 protein level, 48 h after miR-17 ~ 92 transfection in HEK293T, SiHa, HeLa and C33A cell lines compared to vector control. The experiments were done in triplicates. Error bar depicts standard error (S.E.). *p value < 0.05 and **p value < 0.001

Article Snippet: C33A (HPV negative cervical cancer cell line) and HEK293T (Human Embryonic Kidney cell line) were procured from National Centre for Cell Science (NCCS, Pune, India).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Plasmid Preparation

Journal: Discover. Oncology

Article Title: miR-17 ~ 92 suppresses proliferation and invasion of cervical cancer cells by inhibiting cell cycle regulator Cdt2

doi: 10.1007/s12672-023-00775-3

Figure Lengend Snippet: miR-17 ~ 92 was ectopically expressed and the proliferation was observed from day of transfection till day 4. A Growth curve of HEK293T cells with its respective control. B Control and treated HEK293T cells after 48 h of transfection under phase contrast microscopy. C Growth curve of SiHa cells along with its respective control D Control and treated SiHa cells under phase contrast microscopy after 48 h of transfection. E Growth curve of HeLa and its respective control. F Control and treated HeLa cells after 48 h of transfection under phase contrast microscopy. G Growth curve of C33A cells and its respective control. H Control and treated C33A cells after 48 h of transfection under phase contrast microscopy. The experiment was done in triplicate. Error bar represents S.D. *p value- < 0.05, **p value < 0.001

Article Snippet: C33A (HPV negative cervical cancer cell line) and HEK293T (Human Embryonic Kidney cell line) were procured from National Centre for Cell Science (NCCS, Pune, India).

Techniques: Transfection, Microscopy

Journal: bioRxiv

Article Title: miR-17∼92 Suppresses Proliferation and Invasion of Cervical Cancer Cells by Inhibiting Cell Cycle Regulator Cdt2

doi: 10.1101/2022.12.30.522306

Figure Lengend Snippet: miR17-92 over expression decreases Cdt2 expression in cervical cancer cells. A . Different miRNAs from miR 17-92 cluster targets Cdt2/DTL according to different miRNA databases. B . RT-qPCR analysis of mRNA level of Cdt2 48h after transfection of miR-17∼92 in HEK293T, C33A and SiHa C . Western blot for analysis of Cdt2 protein expression level 48 hours after transfection of miR-17∼92 in HEK293T, SiHa, HeLa and C33A cell line. D . Quantification of Cdt2 protein level 48h post miR 17-92 transfection in HEK, SiHa, HeLa and C33A compared to vector control. Quantification experiments were done in triplicates. Error bar depicts standard error (S.E.). *p value <0.05

Article Snippet: C33A (HPV negative cervical cancer cell line) and HEK293T (Human kidney cell line) were procured from National Centre for Cell Science (NCCS, Pune, India).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Plasmid Preparation, Control

Journal: bioRxiv

Article Title: miR-17∼92 Suppresses Proliferation and Invasion of Cervical Cancer Cells by Inhibiting Cell Cycle Regulator Cdt2

doi: 10.1101/2022.12.30.522306

Figure Lengend Snippet: miR17-92 suppresses proliferation of cervical cancer cells: miR17-92 ectopically expressed and the proliferation was observed from day of transfection till day 4 A. Growth curve of HEK293T cells with its respective control B. Control and treated HEK293T cells after 48h of transfection under phase contrast microscopy. C. Growth curve of SiHa along with its control D. SiHa cells (control and treated) under phase contrast microscopy after 48h of transfection. E. Growth curve of HeLa and it’s control F. Phase contrast microscopy of treated and control HeLa cells 48h post transfection. G. Growth curve of C33A and its control after transfection F. C33A control and treated cells under phase contrast microscopy after 8h of transfection. The experiment was done in triplicate. Error bar represents S.D. *p value-<0.05, **p value<0.001

Article Snippet: C33A (HPV negative cervical cancer cell line) and HEK293T (Human kidney cell line) were procured from National Centre for Cell Science (NCCS, Pune, India).

Techniques: Transfection, Control, Microscopy

Journal: International journal of oncology

Article Title: Sine oculis homeobox homolog 1 promotes DNA replication and cell proliferation in cervical cancer.

doi: 10.3892/ijo.2014.2510

Figure Lengend Snippet: Figure 1. SIX1 expression is increased by HPV oncoprotein in cervical cancer. (A) Immunohistochemical analysis of SIX1 protein in tissue microarray of human specimens. Representative samples of SIX1 staining are shown at x200 magnification (left). Bar, 100 µm. The immunohistochemical scores (HSCORE) of SIX1 are also shown (right). (B) The expressions of SIX1 in three cervical cancer cell lines were detected by western blot analysis. (C) The expressions of HPV-E6, HPV-E7 and SIX1 in indicated C33a cells were detected by western blot analysis. (D) At 48 h after HPV-E7 siRNA transfection, the expressions of HPV-E7 and SIX1 in cells detected by western blot analysis. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Human cervical cancer cell lines C33a (HPV-negative), Siha and Caski (HPV-positive) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Immunohistochemical staining, Microarray, Staining, Western Blot, Transfection

Journal: International journal of oncology

Article Title: Sine oculis homeobox homolog 1 promotes DNA replication and cell proliferation in cervical cancer.

doi: 10.3892/ijo.2014.2510

Figure Lengend Snippet: Figure 3. SIX1 regulates the expression of genes responsible for DNA replication. (A and B) In C33a-3.1 and C33a-SIX1 cells, SIX1 expression was detected by (A) western blot analysis, and the expression levels of indicated genes were detected by (B) real-time RT-PCR. (C and D) Two independent shRNAs were used to knockdown SIX1 expression in Caski cells. SIX1 expression was detected by (C) western blot analysis and the expression levels of indicated genes were detected by (D) real-time RT-PCR. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Human cervical cancer cell lines C33a (HPV-negative), Siha and Caski (HPV-positive) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Knockdown

Journal: International journal of oncology

Article Title: Interleukin-6-induced epithelial-mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma.

doi: 10.3892/ijo.2014.2422

Figure Lengend Snippet: Figure 3. Expression of IL-6, IL-6R, p-Stat3, E-Cadherin, Vimentin and p-Slug in HeLa and C33A cells. (A) The protein expression levels of endogenous IL-6, IL-6R, p-Stat3 and p-Slug were weakly detected in C33A cells as compared to HeLa cells, but the protein levels of endogenous E-Cadherin and Vimentin were no different in HeLa and C33A cells. (B-E) The expression levels of IL-6R protein and mRNA were significantly increased in IL-6-treated HeLa and C33A cells for 48 h as compared to control, DMSO and IL-6 + curcumin groups (quantitative results). β-actin was used as an internal control. The intensity of bands was quantified using ImageJ software and normalized to β-actin.

Article Snippet: The HeLa cervical cancer cell line (HPV-positive) and the C33A cervical cancer cell line (HPV-negative) were obtained from American Type Culture Collection.

Techniques: Expressing, Control, Software

Journal: International journal of oncology

Article Title: Interleukin-6-induced epithelial-mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma.

doi: 10.3892/ijo.2014.2422

Figure Lengend Snippet: Figure 4. IL-6-induced cell proliferation and cell morphology alteration. (A) HeLa and C33A cells were treated without IL-6 (as a control) and with IL-6 (50 ng/ml), IL-6 + curcumin (10 µM) for 48 h. Cell proliferation was evaluated by immunostaining using Ki67 antibody. (B) Quantitative analysis shows that IL-6-induced HeLa and C33A cell proliferation as compared to control and IL-6 + curcumin group. (C) HeLa cells were treated with IL-6 for 48 h, alteration of cell morphology was observed by immunostaining using IL-6Rα antibody (bottom panel) as compared to control (upper panel). Nuclei were stained with DAPI (original magnification, x400).

Article Snippet: The HeLa cervical cancer cell line (HPV-positive) and the C33A cervical cancer cell line (HPV-negative) were obtained from American Type Culture Collection.

Techniques: Control, Immunostaining, Staining

Journal: International journal of oncology

Article Title: Interleukin-6-induced epithelial-mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma.

doi: 10.3892/ijo.2014.2422

Figure Lengend Snippet: Figure 5. IL-6 treatment induces EMT in HeLa and C33A cells. (A) HeLa cells were treated without IL-6 (as a control) and with IL-6 (50 ng/ml), IL-6 + curcumin (10 µM) for 48 h. Cells were stained with anti-E-Cadherin antibody and rhodamine-conjugated secondary antibody (red) or stained with anti-Vimentin antibody and fluorescein isothiocyanate-conjugated secondary antibody (green). Nuclei were stained with DAPI (blue) (original magnification, x400). (B and C) HeLa cells were treated without IL-6 (as a control) and with DMSO, IL-6 (50 ng/ml) and IL-6 + curcumin (10 µM) for 48 h. Total protein was extracted from control, DMSO and treated cells. The protein levels of E-Cadherin (B, left) and Vimentin (C, left) were analyzed using western blot analysis. Similarly, total RNA was extracted from control and treated cells. The mRNA levels of E-Cadherin (B, right) and Vimentin (C, right) were determined using real-time RT-PCR. Real-time PCR for GADPH served as an internal control. (D) C33A cells were treated without IL-6 (as a control) and with IL-6 (50 ng/ml), IL-6 + curcumin (10 µM) for 48 h. Cells were stained with anti-E-Cadherin antibody and rhodamine-conjugated secondary antibody (red) or stained with anti-Vimentin antibody and fluorescein isothiocyanate-conjugated secondary antibody (green). Nuclei were stained with DAPI (blue) (original magnification, x400). (E and F) C33A cells were treated without IL-6 (as a control) and with DMSO, IL-6 (50 ng/ml) and IL-6 + curcumin (10 µM) for 48 h. Total protein was extracted from control, DMSO and treated C33A cells. The protein levels of E-Cadherin (E, left) and Vimentin (F, left) were analyzed using western blot analysis. Similarly, total RNA was extracted from control and treated cells. The mRNA levels of E-Cadherin (E, right) and Vimentin (F, right) were determined using real-time RT-PCR. Real-time PCR for GADPH served as an internal control. The intensity of bands was quantified using ImageJ software and normalized to β-actin.

Article Snippet: The HeLa cervical cancer cell line (HPV-positive) and the C33A cervical cancer cell line (HPV-negative) were obtained from American Type Culture Collection.

Techniques: Control, Staining, Western Blot, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Software

Journal: International journal of oncology

Article Title: Interleukin-6-induced epithelial-mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma.

doi: 10.3892/ijo.2014.2422

Figure Lengend Snippet: Figure 6. IL-6-induced phosphorylation of Stat3 by treatment with IL-6 in HeLa and C33A cells. (A) HeLa cells were treated with IL-6 (50 ng/ml) and IL-6 + curcumin (10 µM) for 48 h. HeLa cells were stained with anti-p-Stat3 (Y705) antibody and fluorescein isothiocyanate-conjugated secondary antibody (green). Nuclei were stained with DAPI (blue) (original magnification, x400). Quantitative analysis of p-Stat3 expression is markedly higher in IL-6 treated cells as compared to untreated and IL-6 + curcumin (10 µM) treated cells. (B) HeLa cells were stimulated with IL-6 (5, 50 and 200 ng/ml) and without IL-6 for 48 h. Cell lysates were subjected to immunoblot analysis using anti-p-Stat3 antibody and β-actin as loading control. (C) HeLa cells were treated without IL-6 and with DMSO, IL-6 (50 ng/ml) and IL-6 + curcumin (10 µM) for 48 h. Cell lysates were subjected to immunoblot analysis using anti-p-Stat3 and anti-Stat3 antibodies. β-actin was a loading control. (D) C33A cells were treated with IL-6 (50 ng/ml) for 48 h. Cells were stained with anti-p-Stat3 (Y705) antibody and rhodamine-conjugated secondary antibody (red) (original magnification, x400). (E) C33A cells were stimulated with IL-6 (5, 50 and 200 ng/ml) and without IL-6 (as a control) for 48 h. Cell lysates were subjected to immunoblot analysis using anti-p-Stat3 antibody and β-actin as a loading control. (F) C33A cells were treated without IL-6 (as a control) and with DMSO, IL-6 (50 ng/ml) and IL-6 + curcumin (10 µM) for 48 h. Cell lysates were subjected to immunoblot analysis using anti-p-Stat3 and anti-Stat3 antibodies. β-actin was used as a loading control. The intensity of bands was quantified using ImageJ software and normalized to β-actin.

Article Snippet: The HeLa cervical cancer cell line (HPV-positive) and the C33A cervical cancer cell line (HPV-negative) were obtained from American Type Culture Collection.

Techniques: Phospho-proteomics, Staining, Expressing, Western Blot, Control, Software

Journal: International journal of oncology

Article Title: Interleukin-6-induced epithelial-mesenchymal transition through signal transducer and activator of transcription 3 in human cervical carcinoma.

doi: 10.3892/ijo.2014.2422

Figure Lengend Snippet: Figure 7. Stat3 knockdown significantly reverses the IL-6-induced EMT program in HeLa and C33A cells. (A) Stat3 was knocked down in HeLa cells by respective siRNA. Cells treated with or without IL-6 for 48 h and whole-cell extracts were analyzed by SDS-PAGE. The protein expression levels of p-Stat3, E-Cadherin, Vimentin and p-Slug were examined by western blot analysis. Equal protein loading was verified by stripping the blots and reprobing with β-actin antibody. (B) Stat3 was knocked down in C33A cells by respective siRNA. Cells were treated with or without IL-6 for 48 h, and cell lysates were subjected to immunoblot analysis using anti-p-Stat3, anti-E-Cadherin, anti-Vimentin and anti-p-Slug antibodies. The intensity of bands was quantified using ImageJ software and normalized to β-actin as a loading control.

Article Snippet: The HeLa cervical cancer cell line (HPV-positive) and the C33A cervical cancer cell line (HPV-negative) were obtained from American Type Culture Collection.

Techniques: Knockdown, SDS Page, Expressing, Western Blot, Stripping Membranes, Software, Control