Two-way analysis of variance (ANOVA) followed by Tukeys multiple comparisons test was performed to compare significant difference and calculate the test and Two-way ANOVA followed by Tukeys multiple comparisons test, *test and Two-way ANOVA followed by Tukeys multiple comparisons test, *test, *test, **respectively. Additional file Porcn-IN-1 7: Figure S4. Porcn-IN-1 Knockdown of COL1A1 inhibits the proliferation and migration of HCC cells. 13046_2020_1650_MOESM7_ESM.tif (3.9M) GUID:?37FABE13-FD92-42FC-9991-2937853EB5F1 Additional file 8: Figure S5. Knockdown of COL3A1 has no effect on cell proliferation and migration. 13046_2020_1650_MOESM8_ESM.tif (4.6M) GUID:?A890CE40-173F-40BC-9023-9902C2F3B481 Additional file 9: Figure S6. RUNX1 is a transcriptional factor of COL4A1. 13046_2020_1650_MOESM9_ESM.tif (7.0M) GUID:?F2AF57F3-EA37-4050-AF5B-31487825CAA5 Additional file 10: Figure S7. Collagen IV activates the FAK-Src signaling. 13046_2020_1650_MOESM10_ESM.tif (4.8M) GUID:?8DA4E622-9EAD-4AE5-B406-5A7F9113B74D Data Availability StatementThe data supporting our conclusion were obtained from the TCGA database Porcn-IN-1 (https://cancergenome.nih.gov), Oncomine database (https://www.oncomine.org), GEO datasets (https://www.ncbi.nlm.nih.gov/gds/), and Human Protein Atlas online database (https://www.proteinatlas.org). Abstract Background Collagens are the most abundant proteins in extra cellular matrix and important components of tumor microenvironment. Recent studies have showed that aberrant expression of collagens can influence tumor cell behaviors. However, their roles in hepatocellular carcinoma (HCC) are poorly understood. Methods In this study, we screened all 44 collagen members in HCC using whole transcriptome sequencing data from the public datasets, and collagen type IV alpha1 chain (COL4A1) was identified as most significantly differential expressed gene. Expression of COL4A1 was detected in HCC samples by quantitative real-time polymerase chain reaction (qRT-PCR), western blot and immunohistochemistry (IHC). Finally, functions and potential mechanisms of COL4A1 were explored in HCC progression. Results COL4A1 is the most significantly overexpressed collagen gene in HCC. Upregulation of COL4A1 facilitates the proliferation, migration and invasion of HCC cells through FAK-Src signaling. Expression of COL4A1 is upregulated by RUNX1 in HCC. HCC cells with high COL4A1 expression are sensitive to the treatment with FAK or Src inhibitor. Conclusion COL4A1 facilitates growth and metastasis in HCC via activation of FAK-Src signaling. High level of COL4A1 may be a potential biomarker for diagnosis and treatment with FAK or Src inhibitor for HCC. test (paired/unpaired). Pearson correlation Porcn-IN-1 tests were performed on correlation analyses. Two-way analysis of variance (ANOVA) followed by Tukeys multiple comparisons test was performed to compare significant difference and calculate the test and Two-way ANOVA followed by Tukeys multiple comparisons test, *test and Two-way ANOVA followed by Tukeys multiple comparisons test, *test, *test, **respectively. Data are presented as means standard deviation. Student test, * em P /em ? ?0.05, ** em P /em ? ?0.01, *** em P /em ? ?0.001, **** em P /em ? ?0.0001, ns, not significant Discussion In this study, we first put forward the role of COL4A1 in HCC tumorigenesis. COL4A1 is dramatically upregulated collagen gene in HCC by screening the expression patterns of all 44 collagen genes in liver cancer from the TCGA-LIHC database. COL4A1 promotes the growth and metastasis of HCC cells by activating FAK-Src signaling. RUNX1 is a transcriptional factor of COL4A1 and activates the expression of COL4A1 in HCC. Targeting FAK or Src may be an effective strategy Ceacam1 to treat HCC patients with high expression of COL4A1 (Fig. ?(Fig.77). Open in a separate window Fig. 7 Schematic diagram of COL4A1 promoting the growth and metastasis of HCC cells. COL4A1 promotes the growth, migration and invasion of HCC cells by activating FAK-Src signaling. Col IV, Collagen type IV Collagen proteins form the scaffold of tumor microenvironment and are important for tumor infiltration, angiogenesis, and metastasis [5]. Some collagen genes have been found aberrant expression during carcinogenesis in various types of cancer. However, only a few studies on the expression and function of collagen genes have been reported in HCC. Some studies reported that COL1A1 was upregulated in HCC and could promote HCC progression [17, 40, 41]. Based on bioinformatics analysis, Liu et al. reported that COL4A1 and COL4A2 were significantly correlated with hepatocarcinogenesis and HCC progression [56]. In this study, we analyzed the expression patterns of all 44 collagen genes in liver cancer from TCGA-LIHC database, and found that the expression of around 70% collagen genes are dysregulated. Among these dysregulated collagen genes, expression of COL4A1 is most abundant and significantly upregulated in HCC. Although Col IV has been reported to associate with the progression of cancer [36, 46], the fine detail molecular mechanisms aren’t well recorded. Burnier et al. demonstrated that Col IV triggered FAK in liver organ metastasis sites produced by different major tumors [57]. Our data demonstrated COL4A1 manifestation could influence the phosphorylation of FAK in HCC cells, recommending that COL4A1 activates FAK signaling to market HCC development. Chen et al. demonstrated that COL4A1 controlled tumor cell migration and stiffness through activation of Src and ERK1/2 [46]. Espinosa et al. reported that Col IV improved the activation and expression of ERK1/2 [53]. In breast tumor, COL4A1 induced MMP-9 manifestation by activating Src phosphorylation [54]. Inside our research, COL4A1 overexpression improved the phosphorylation of Src, but had simply no effect on manifestation degree of phosphorylation and MMP-9 of ERK1/2 in HCC.