Introduction

Anthocyanins decrease the proliferation of colorectal cancer cell and cholesterol regulation via the adenosine triphosphate-binding cassette subfamily A member 1 (ABCA1) protein, which is underexpressed in colon cancer.^1,2,3 Anthocyanins, which are found in Indonesian native black rice bran, can also promote the outflow of cholesterol from cancer cell.

Aims & Methods

This study aims to explore the influence of Anthocyanins Black Rice Bran Extract (ABRiBE) on mRNA expression of the ABCA1 gene and colorectal cancer cell proliferation in the human colorectal cancer cell line, HT-29. ABRiBE is thought to suppress the proliferation of HT-29 cell while increasing the expression of the ABCA1 gene. Anthocyanins derived from black rice bran powder using ethanol acidified with citric acid were applied in this experimental work on HT-29 cells. The MTS assay used 12 extract concentrations and control groups to determine the inhibitory concentration (IC₅₀) of 50% cell growth inhibition for 24 and 48 hours. The quantitative Polymerase Chain Reaction (qPCR) method was used to examine the expression of the ABCA1 gene in groups higher than the IC₅₀ value and a control group.

Results

The proliferation of HT-29 cells was lowered by ABRiBE, which had an IC₅₀ of 1.57 and 1.35 mg/mL for incubation of 24 and 48 hours with values of r=0.97, p=0.001 and r=0.95, p=0.001, respectively. The expression of the ABCA1 gene was enhanced by 1.16 and 2.32 fold when extract supplementating with 2 doses beyond the IC₅₀ values of 1.7 and 2.0 mg/mL and a control group.

Table 1. The mRNA relative expression of the ABCA1 and HRPT1 (reference) genes as well as their primer pair sequence and amplicon sizes

Conclusion

The ABRiBE decreased the proliferation of the HT-29 cells while increasing the expression of the ABCA1 gene.

References

1. Mazewski C, Liang K and Gonzalez de Mejia E. 2018. Comparison of the effect of chemical composition of anthocyanin rich plant extracts on colon cancer cell proliferation and their potential mechanism of action using in vitro, in silico, and biochemical assays. Food Chem., 242:378-388. doi:10.1016/j.foodchem.2017.09.086
2. Smith B and Land H. 2012. Anticancer activity of the cholesterol exporter ABCA1 gene. Cell Rep., 2(3):580-590. doi:10.1016/j.celrep.2012.08.011
3. Zhao X, Feng P, He W, Du X, Chen C, Suo L, Liang M, Zhang N, Na A and Zhang Y. 2019. The prevention and inhibition effect of anthocyanins on colorectal cancer. Curr Pharm Des., 25(46):4919-4927. doi:10.2174/1381612825666191212105145

Disclosure

The authors declare no conflict of interest.

Introduction

One of major obstacles of treatment in colorectal cancer (CRC) is drug resistance, specially 5-Fu chemoresistance which is a fundamental drug in CRC chemotherapy. Also, there is no reliable biomarkers in order to identify chemoresistance patients and improve their treatment. There are several factors that cause 5-FU chemoresistance include autophagy. This cell death phenomenon is played key role in 5-FU chemoresistance in CRC patients by providing nutritarians for CRC cells in hard circumstances; also, autophagy has regulated by different factors like genes and non-coding RNAs(ncRNA).

Aims & Methods

Verifying key gene in autophagy pathway at KEGG database. Moreover, evaluation the expression of key gene in CRC patients at TCGA database. Investigation on the key gene related LncRNAs and miRNAs in CRC at LNCpedia and mirTARBase database respectively, in order to find most relevant ncRNAs of this gene and obtain proper biomarkers. Developing 5-FU chemoresistance Caco2 and SW480 cell lines by increasing dosage of drug and MTT assay was performed to confirm the chemoresistance cell lines and western blot was performed to confirm autophagy in cell lines. Finally, evaluating the expression of MAP1LC3A and its related ncRNAs in 5-FU chemoresistant CRC cell lines by qRT-PCR.

Results

MAP1LC3A was confirmed as key gene based on KEGG analysis; also, FAM225A, NCK1-DT, LINC01825, LINC02188 and miR-335-5p have nominated as most relevant ncRNAs based on bioinformatic analysis. Furthermore, qRT-PCR analysis of MAP1LC3A and its related ncRNAs have shown differential expression between chemoresistant and chemoresistant CRC cell lines.

Conclusion

These findings indicate that FAM225A, NCK1-DT, LINC01825, LINC02188 and miR-335-5p participate in autophagy by effecting the expression of MAP1LC3A gene and they can function as proper biomarkers for 5-FU chemoresistance CRC patients. Not only they can function as biomarkers but also, they can attend in target therapy by designing siRNAs to knockdown this ncRNAs and enhance chemotherapy in CRC patients.

Introduction

Colorectal cancer (CRC) is one of the most common digestive tract malignant tumors, and its incidence rate and mortality rate rank third among all malignant tumors. The IMMUNOGLOBULIN SUPERFAMILY DOMAINS genes has been reported to be hypermethylated in human colorectal cancer (CRC) cell lines and primary tumor tissues, .On the other hand High mobility group genes are characteristically present at high levels in a variety of human cancers including colorectal, ovarian, breast, lung, and pancreatic cancer.

Aims & Methods

The aim of the work to evaluate the gene expression of both leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) (as an example of immunoglobulin sperfamily domains and High mobility group AT-hook 2 (HMGA2) as an example of high mobility group as diagnostic and prognostic markers in cases of colorectal cancer.
In this study forty-five patients pathologically approved with colorectal cancer were prospectively analyzed for LRIG1 and HMGA2 genes expression at tissue level. SYBR Green Real-Time RT-PCR technique was used for gene expression analyses and β-actin gene was considered as a reference gene for data normalization. Additionally, appropriate statistical analyses were used to assess the expression of LRIG1 and HMGA2 in diseased and adjacent healthy tissues.

Results

HMGA2 mRNA gene expression was over-expressed in CRC tissue than normal tissue ( median fold change and median Log² ratio are [9.3(0.2, 100.4) and 3.1(-2.6, 6.7), respectively]with positive correlation between HMGA2 and poor differentiation ,more advanced stage and lymph node metastasis(p> 0.05) but not with distant metastasis (p >0.05) , The expression of LRIG1 mRNA gene in tumor tissue was low expressed in colorectal tumor tissues (Median fold change and median Log2 ratio are [0.29(0.01, 4.14) and -1.8(-6.1, 2.1), respectively but no correlation of the LRIG1 gene with the the stage, Differentiation and metastasis of the CRC (P >0.05). There is no correlation of both genes neither with tumor biomarkers (CA19-9, CEA) nor with laboratory findings (WBCs, Hemoglobin, Platelet, INR, Albumin, Creatinine) P>0.5%). The overall survival showed no significant association with any genetic expression of both genes (P >0.05).

Conclusion

LRIG1 low expression may be considered for diagnosis but not valuable as prognostic marker in colorectal cancer patients. However, HMGA2 overexpression may be valuable as both diagnostic and prognostic markers in CRC.

Disclosure

No conflict of interest to be disclosed

Introduction

As common precursors to colorectal cancer (CRC), polyps and adenomas in the large intestine may develop into malignancy over time through a cascade known as the "normal-polyp-adenoma-carcinoma" sequence. Colorectal cancer (CRC) is the third most common cancer and the leading cause of cancer-related death worldwide^​, with an estimated increase to 3.2 million new cases and 1.6 million deaths by 2040^​. The high mortality of CRC is largely due to the late diagnosis, uncomplete molecular understanding of pathological progression, and limited treatment options for patients with advanced disease stage. Therefore, the identification of novel biomarkers at early (pre-cancerous) phase, thorough elucidation of pathological mechanisms, and development of effective therapies is crucial for CRC diagnosis and treatment.

Aims & Methods

Here we have utilized 10× genomics scRNA-seq platform to perform a high-resolution transcriptomic analysis of three normal colorectal tissues, two samples with inflammatory polyps, two specimens with adenomatous polyps and three CRC samples.

Results

By analyzing the scRNA-seq data of the ten human colorectal samples, which represent the four phases of colorectal pathogenesis, in combination with validation by immunofluorescence staining using expanded clinical samples, and together with verification using large scale TCGA and Kaplan–Meier data, we revealed a dynamic molecular landscape and for the first time demonstrated the importance of ligand-receptor genes, transcription factors and genes encoding multiple immunoglobulins and heat shock proteins driving the colorectal carcinogenesis.
Besides the heterogeneity of (sub-) cell types and difference of the transcriptomic profiling across the four stages, one of the most interesting and novel findings of this study is that the expression of ligand-receptor genes can be used to determine the cell types and even cell sub-types, at least for the colorectal tissues collected in this study, not the expressions of other gene sets, such as cell surface receptor signaling pathways, cell-cell signaling, transcription factors, cytokines, cell activation factors, cell signaling factors, or extracellular matrisome, implying that ligand-receptor pairs are fundamentally important for the cell type determination and cell-cell communications in colorectal tissues.
We found that some genes encoding immunoglobulin members are highly expressed in the T cells (IGHA1, IGHA2, IGKC and IGLC2) and in the fibroblasts (IGHA1, IGHA2, IGLC1, IGLC2, IGKC and JCHAIN) from normal colorectal tissues, while some genes encoding heat shock proteins are highly expressed in T cells (HSP90AA1, HSP90AB1, HSPA1B, HSPD1, HSPE1 and HSPH1) and plasma cells (HSP90AA1, HSPA1A, HSPA1B, HSPA6, HSPB1 and HSPD1) from abnormal colorectal samples, especially in the ones with adenomatous polyps.
Furthermore, IgG plasma cells can be grouped into two sub-types, characteristic of high expression of immunoglobulin-encoding genes IGKC and IGLC1/IGLC2, respectively. In addition, the six clusters of IgA plasma cells can also be characterized based on the combinatory expression level of some immunoglobulin-encoding genes (IGHA1, IGHA2, IGKC, IGLC1, IGLC2 and IGLC3). In summary, our analyses suggested the potential distinct functional importance of immunoglobulins and heat shock proteins in immunity and microenvironment during the colorectal malignancy.

Conclusion

Our transcriptome data provides valuable insights and a useful resource for better understanding the molecular pathological landscape in CRC to achieve more effective diagnosis and treatments.

References

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