Related Experiment Video
Updated: Jun 2, 2026
An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis
Published on: September 12, 2019
[Effect of RhoC on hepatocellular carcinoma cell growth and related molecular mechanisms]
Shu-li Xie1, Ming-guang Zhu, Guo-yue Lü
1Department of General Surgery, the First Hospital of Jilin University, Changchun 130021, China. xieshuli6310@hotmail.com
Objective:
To clarify the role of RhoC in the growth of hepatocellular carcinoma cells and its molecular mechanism, so as to explore the molecular target of tumor cell growth.
Methods:
siRNA-RhoC plasmid was constructed and RhoC gene silencing the cell-line of hepatocellular carcinoma was setup. Cell growth was assessed by MTT assay. AgNORs staining was applied to determine cell proliferation. Plate cell clone test was conducted to examine the capacity of cell clone formation. FACS was adopted to measure the course of cell cycle and semi-quantitative RT-PCR was used to determine the expression of cell cycle proteins. In order to further determine the effect of RhoC expression on cell growth, a RhoC over-expression human hepatocellular cell line was setup by PcDNA3-RhoC plasmid transfection.
Results:
The inhibition rate of RhoC was 82.3%. From the fourth day of cell culture, the growth of cells in RNAi group was significantly slower than that in parental Bel7402 and negative control groups (0.41 ± 0.10 vs. 0.73 ± 0.11 and 0.71 ± 0.07 respectively, P < 0.05). AgNORs staining showed that average cell stained particles in RNAi group was significantly lower than that in parental Bel7402 and negative control(1.23 ± 0.35 vs. 3.47 ± 0.93 and 3.17 ± 0.78, P < 0.01). Plate clone formation test showed that clone formation efficiency in the RNAi group was notably lower than that in the control group [(20.33 ± 5.42)% vs. (70.58 ± 10.10)% and (69.83 ± 14.77)%, respectively, P < 0.01]. Cell cycle analysis by FACS showed that G(0)/G(1) cell percentage in the RNAi group was significantly higher than that in the control group [(73.14 ± 5.93)% vs. (57.05 ± 5.97)% and (52.99 ± 4.80)%, P < 0.05]. Compared with Bel7402 and negative control groups, the expression of following growth associated genes was significantly decreased: cyclin D1(0.45 ± 0.21 vs. 1.25 ± 0.24 and 1.12 ± 0.15, respectively, P < 0.05)and CDK4 (0.55 ± 0.08 vs. 1.18 ± 0.32 and 1.10 ± 0.29, respectively, P < 0.05); the following genes were notably increased: p16(1.07 ± 0.23 vs. 0.36 ± 0.12 and 0.35 ± 0.13, respectively, P < 0.01)and p21(0.42 ± 0.12 vs. 0.17 ± 0.06 and 0.19 ± 0.08, respectively, P < 0.05). RhoC was highly expressed in PcDNA3-RhoC transfected hepatocellular cell line. From the third day on of the cell culture, cell growth in PcDNA3-RhoC group was remarkably higher than that in the HL7702 and PcDNA3 groups (0.83 ± 0.10 vs. 0.54 ± 0.11 and 0.58 ± 0.55, respectively, P < 0.05).
Conclusions:
RhoC is the key molecule in promoting hepatocellular cell growth, and is a promising target for tumor cell growth controlling.
Insights
RhoC protein is crucial for hepatocellular carcinoma cell growth. Silencing RhoC significantly inhibits tumor cell proliferation and progression, identifying it as a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Understanding the molecular mechanisms driving HCC growth is critical for developing effective therapies.
Purpose of the Study:
- To investigate the role of RhoC in hepatocellular carcinoma cell proliferation.
- To elucidate the molecular mechanisms underlying RhoC's function in HCC growth.
- To identify RhoC as a potential therapeutic target for HCC.
Main Methods:
- RhoC gene silencing using siRNA in HCC cell lines.
- Cell proliferation assessed by MTT assay and AgNORs staining.
- Clonogenic potential evaluated through plate clone formation tests.
- Cell cycle analysis using Flow Cytometry (FACS).
- Expression of cell cycle regulatory proteins determined by semi-quantitative RT-PCR.
- RhoC overexpression model established for further validation.
Main Results:
- RhoC silencing led to an 82.3% inhibition rate, significantly reducing cell growth, proliferation, and colony formation.
- FACS analysis revealed an increased G0/G1 phase cell population upon RhoC silencing.
- Downregulation of cyclin D1 and CDK4, and upregulation of p16 and p21 were observed post-RhoC silencing.
- RhoC overexpression significantly enhanced hepatocellular cell growth.
Conclusions:
- RhoC plays a pivotal role in promoting hepatocellular carcinoma cell growth.
- Targeting RhoC represents a promising strategy for controlling HCC tumor progression.
