hcc cell line hep 3b  (ATCC)

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: Identification of circSP3 in HCC cells and confirmation of its circular structure. ( A ) The predicted binding between circSP3 and miR-198. ( B ) Schematic illustration demonstrating that circSP3 is located on chromosome 2q31.1 and cyclized from exons of SP3 . ( C ) The PCR products of circSP3 were confirmed through Sanger sequencing. ( D ) CircSP3 was detected in Huh-7 cells. As expected, divergent primers detected circSP3 in cDNA but not in gDNA. GAPDH was used as a negative control. ( E ) The relative levels of circSP3 and SP3 were assessed through qRT-PCR analysis in cells treated with or without RNase R. *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Binding Assay, Sequencing, Negative Control, Quantitative RT-PCR

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: Expression of circSP3 in HCC tissues and cell lines. ( A ) CircSP3 levels in 48 pairs of HCC tissues and adjacent normal liver tissues were evaluated using qRT-PCR. ( B ) The relative circSP3 levels in four HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and an immortalized liver cell line (HL-77O2) were determined using qRT-PCR. ( C ) CircSP3 levels in Hep-3B cells infected with NC or circSP3 plasmids. ( D ) qRT-PCR was conducted to confirm the knockdown efficiency of si-circSP3-1 and si-circSP3-2. ( E ) qRT-PCR was conducted to confirm the knockdown efficiency of LV3-circSP3. *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Infection, Knockdown

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: CircSP3 induced HCC cell proliferation in vitro . ( A ) A CCK-8 assay was performed to assess cell proliferation when circSP3 was overexpressed in Hep-3B cells. ( B ) A CCK-8 assay was performed to assess cell proliferation when circSP3 was knocked down in Huh-7 cells. ( C ) The cell cycle was analyzed using flow cytometry when circSP3 was overexpressed or knocked down. *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, CCK-8 Assay, Flow Cytometry

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: CircSP3 induced HCC cell migration and invasion in vitro . ( A , B ) Cell migration and invasion were assessed with Transwell assays in Hep-3B cells infected with circSP3 or NC plasmids. ( C , D ). Transwell assays were carried out to assess cell migration and invasion when circSP3 was downregulated in Huh-7 cells. **p<0.01.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, In Vitro, Infection

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: The relationship between circSP3 and miR-198 in HCC. ( A ) The relative miR-198 levels in four HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were assessed using qRT-PCR. ( B ) Correlation analysis between circSP3 and miR-198 levels in HCC tissues. ( C ) The relative miR-198 levels in Hep-3B cells infected with NC or circSP3 plasmids were detected using qRT-PCR. ( D ) The relative miR-198 levels in Huh-7 cells infected with si-NC or si-circSP3 plasmids were detected using qRT-PCR. ( E ) qRT-PCR revealed that ectopic expression of miR-198 reduced circSP3 expression in Huh-7 cells. ( F ) qRT-PCR demonstrated that knocking down miR-198 increased circSP3 expression in Hep-3B cells. ( G ) Schematic of predicted wild-type and mutated miR-198 binding sequences in circSP3. ( H , I ) Luciferase activity in HCC cells co-transfected with circSP3-wt or circSP3-mut and miR-198 mimics or miR-198 mimic controls. ( J ) FISH revealed that miR-198 colocalized with circSP3 in HCC cells. **p<0.01, ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Quantitative RT-PCR, Infection, Expressing, Binding Assay, Luciferase, Activity Assay, Transfection

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: CircSP3 induces HCC cell proliferation, migration and invasion by sponging miR-198 and upregulating CDK4. CircSP3-overexpressing Hep-3B cells were infected with miR-198 mimics or miR-198 mimic controls, and circSP3-knockdown Huh-7 cells were infected with miR-198 inhibitors or miR-198 inhibitor controls. ( A , B ) CCK-8, ( C , D ) Transwell migration and ( E , F ) Transwell invasion assays were conducted to assess cell proliferation, migration and invasion, respectively. ( G , H ) The effects of circSP3 and miR-198 expression on CDK4 expression were quantified using Western blotting. *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, Infection, Knockdown, CCK-8 Assay, Expressing, Western Blot

Journal: Aging (Albany NY)

Article Title: Circular RNA circSP3 promotes hepatocellular carcinoma growth by sponging microRNA-198 and upregulating cyclin-dependent kinase 4

doi: 10.18632/aging.203303

Figure Lengend Snippet: CircSP3 promotes HCC cell proliferation, migration and invasion by inducing CDK4. CircSP3-overexpressing Hep-3B cells were infected with CDK4-homo-520 ( CDK4 knockdown) or CDK4-homo-520 control plasmids, and circSP3-knockdown Huh-7 cells were infected with CDK4-overexpressing or CDK4 control plasmids. ( A , B ) CCK-8, ( C , D ) Transwell migration and ( E , F ) Transwell invasion assays were conducted to assess cell proliferation, migration and invasion, respectively. ( G , H ) The effects of circSP3 and CDK4 expression plasmids on CDK4 expression were quantified using Western blotting. *p<0.05, **p<0.01 and ***p<0.001.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Bel-7402 and SMMC-7721) and a normal liver cell line (HL-77O2) were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, Infection, Knockdown, Control, CCK-8 Assay, Expressing, Western Blot

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ_0003418 downregulation in HCC tissues and cells. Notes: ( A ) The expression profile of circ_0003418 in HCC tissues and adjacent noncancerous tissues were detected by qRT-PCR assays. ( B ) qRT-PCR was used to analyze the level of circ_0003418 in five HCC cell lines and one normal human hepatocyte cell line. ( C and D ) The knockdown efficiency of LV3-circ_0003418 on circ_0003418 in Huh-7 and Hep-3B cells was verified via qRT-PCR. ** P<0.01 and *** P<0.001 compared to control group. Abbreviations: HCC, hepatocellular carcinoma; NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ_0003418 suppresses proliferation, migration, and invasion and promotes apoptosis in HCC cells. Notes: ( A and B ) CCK-8 assays were performed to measure the effect of silencing circ_0003418 on the proliferation in Huh-7 and Hep-3B cells. ( C–F ) Effect of silencing circ_0003418 on cell migration ( C and D ) and invasion ( E and F ) were analyzed by transwell migration and invasion assays, respectively. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviations: CCK-8, cell counting kit 8; NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Migration, CCK-8 Assay, Control, Cell Counting, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ-0003418 sensitizes HCC cells to cisplatin in vitro. Notes: ( A and B ) Huh-7 and Hep-3B cells infected with LV3-NC or LV3-circ_0003418 were treated with different doses of cisplatin (1, 2, 4, 8, 16, 32, 64 and 128 mg/L) for 24 hrs, and then cell viability was determined by CCK-8 assays. ( C–F ) Huh-7 cells were treated with cisplatin (11.39 mg/L) for 24 hrs as well as Hep-3B cells were treated with cisplatin (20.18 mg/L) for 24 hrs, and then cell migration and invasion were detected by transwell migration ( C and D ) and invasion ( E and F ) assays, respectively. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vitro, Infection, CCK-8 Assay, Migration, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: circ-0003418 enhances sensitivity of HCC cells to cisplatin in vivo. Female BALB/c nude mice were implanted subcutaneously with Huh-7 cells infected with LV3-NC or LV3-circ_0003418. Twelve days later, the LV3-NC tumor-bearing mice were treated with saline or cisplatin (5 mg/kg) by intraperitoneal injection twice a week up to 36 days posttreatment. The mice bearing LV3-circ_0003418 tumors received the same treatment. Notes: ( A ) Image of the tumors in the nude mice. ( B ) The tumors growth curve of xenograft model mice. ( C and D ) The weight and volume of the tumors in the nude mice. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: In Vivo, Infection, Saline, Injection, Control, Negative Control

Journal: OncoTargets and therapy

Article Title: circ_0003418 Inhibits Tumorigenesis And Cisplatin Chemoresistance Through Wnt/β-Catenin Pathway In Hepatocellular Carcinoma

doi: 10.2147/OTT.S229507

Figure Lengend Snippet: Silencing circ_0003418 induces cisplatin resistance of HCC cells through activating the Wnt/β-catenin pathway. Notes: ( A and B ) Huh-7 cells were treated with cisplatin (11.39 mg/L) for 24 hrs as well as Hep-3B cells were treated with cisplatin (20.18 mg/L) for 24 hrs, and then the protein levels of β-catenin and c-Myc in the Huh-7 and Hep-3B cells were detected by Western blotting. ( C and D ) The inhibition efficiency of ICG-001 was detected by Western blotting. ( E and F ) Cell proliferation assay showed that inhibition of Wnt/β-catenin pathway in cells infected with LV3-circ_0003418 inhibited cell proliferation. ( G and H ) Chemotherapy sensitivity assay showed that inhibition of Wnt/β-catenin pathway in cells infected with LV3-circ_0003418 enhanced sensitivity of HCC cells to cisplatin. * P<0.05, ** P<0.01 and *** P<0.001 compared to control group. Abbreviation: NC, negative control.

Article Snippet: Human HCC cell lines (Hep-3B, Huh-7, Sk-hep-1, SMMC-7721 and PLC) and normal human hepatocyte line (HL-77O2) were purchased from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Western Blot, Inhibition, Proliferation Assay, Infection, Sensitive Assay, Control, Negative Control