Homeobox A7 promotes esophageal squamous cell carcinoma progression through C-C motif chemokine ligand 2-mediated tumor-associated macrophage recruitment.

Homeobox A7 (HOXA7) plays essential roles in multiple malignancies and was reported to be overexpressed in esophageal squamous cell carcinoma (ESCC). However, its functions in the ESCC tumor microenvironment remain to be explored. In this study, we showed that HOXA7 was overexpressed in ESCC among HOXA family members and correlated with tumor-associated macrophage (TAM) infiltration both in The Cancer Genome Atlas database and ESCC clinical samples. Moreover, transactivation of C-C motif chemokine ligand 2 (CCL2) by HOXA7 was identified (real-time quantitative PCR [RT-qPCR], western blot analysis, ELISA, and ChIP-qPCR), which was detected to drive chemotaxis and M2 polarization of macrophages both in vitro (Transwell assay) and in vivo (xenograft tumors models). In addition, CCL2 triggers macrophage expression of epidermal growth factor (EGF) (RT-qPCR and ELISA), which promotes tumor proliferation and metastasis by activating its receptor EGFR. In addition, EGF-induced ESCC cell proliferation and migration can be abrogated by HOXA7 knockdown (CCK-8 proliferation assay, EdU fluorescence, and Transwell assay). These results indicate a novel mechanistic role of HOXA7 in the cross-talk between ESCC and TAMs, which could be an underlying therapeutic target for ESCC.

Long noncoding RNA MyD88 functions as a promising diagnostic biomarker in hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is one of the most frequent malignancies. Alpha-fetoprotein (AFP) has some limitations in diagnosing early HCC. Recently, long noncoding RNAs (lncRNAs) showed great potential as tumor diagnostic biomarkers, and lnc-MyD88 was previously identified as a carcinogen in HCC. Here, we explored its diagnostic value as a plasma biomarker. Materials and methods: Quantitative real-time PCR was adopted to detect lnc-MyD88 expression in plasma samples of 98 HCC patients, 52 liver cirrhosis (LC) patients, and 105 healthy people. The correlation between lnc-MyD88 and clinicopathological factors was analyzed through chi-square test. The receiver operating characteristic (ROC) curve was used to analyze the sensitivity, specificity, Youden index, and area under the curve (AUC) of lnc-MyD88 and AFP alone and in combination for the diagnosis of HCC. The relationship between MyD88 and immune infiltration was analyzed by single sample gene set enrichment analysis (ssGSEA) algorithm. Results: Lnc-MyD88 was highly expressed in plasma samples of HCC and hepatitis B virus (HBV)-associated HCC patients. Lnc-MyD88 had better diagnostic value than AFP in HCC patients using healthy people or LC patients as control (healthy people, AUC: 0.776 vs. 0.725; LC patients, AUC: 0.753 vs. 0.727). The multivariate analysis showed that lnc-MyD88 had great diagnostic value for distinguishing HCC from LC and healthy people. Lnc-MyD88 had no correlation with AFP. Lnc-MyD88 and AFP were independent diagnostic factors for HBV-associated HCC. The AUC, sensitivity, and Youden index of the combined diagnosis of lnc-MyD88 and AFP combined were higher than those of lnc-MyD88 and AFP alone. The ROC curve of lnc-MyD88 for the diagnosis of AFP-negative HCC was plotted with a sensitivity of 80.95%, a specificity of 79.59%, and an AUC value of 0.812 using healthy people as control. The ROC curve also presented its great diagnostic value using LC patients as control (sensitivity: 76.19%, specificity: 69.05%, AUC value: 0.769). Lnc-MyD88 expression was correlated with microvascular invasion in HBV-associated HCC patients. MyD88 was positively correlated with infiltrating immune cells and immune-related genes. Conclusion: The high expression of plasma lnc-MyD88 in HCC is distinct and could be utilized as a promising diagnostic biomarker. Lnc-MyD88 had great diagnostic value for HBV-associated HCC and AFP-negative HCC, and it had higher efficacy in combination with AFP.

Rs3746444 T>C locus in miR-499 increases the susceptibility to hepatocellular carcinoma: A meta-analysis 14812 subjects.

BACKGROUND: Recently, many investigations have suggested that the rs3746444 T>C locus in the microRNA (miR)-499 gene may contribute to the occurrence of cancer. However, reports on the association between rs3746444 and hepatocellular carcinoma (HCC) are conflicting. AIM: To further understand and explore the potential correlation between the single-nucleotide polymorphism of rs3746444 and the incidence of HCC. METHODS: In this meta-analysis, we obtained electronic literature by searching the PubMed, Embase and Chinese BioMedical Disc databases (through May 20, 2022). All eligible case-control, prospective cohort or nested case-control studies with sufficient data for calculating the odds ratios with their 95% confidence intervals were included. RESULTS: Ultimately, a total of 17 independent studies were included. We identified that rs3746444 was associated with the development of HCC (C vs T: P = 0.019 and CC/CT vs TT: P = 0.016). In Asian individuals, rs3746444 was associated with susceptibility to HCC (C vs T: P = 0.013 and CC/CT vs TT: P = 0.016). In addition, this study identified that the miR-499 rs3746444 locus was associated with susceptibility to HCC in the normal/healthy control subgroup (C vs T: P = 0.034 and CC/CT vs TT: P = 0.024). CONCLUSION: In summary, this meta-analysis highlights that rs3746444 in the miR-499 gene is involved in the occurrence of HCC, especially in Asian individuals.

Lack of association between miR-146a rs2910164 C/G locus and colorectal cancer: from a case-control study to a meta-analysis.

Previous studies suggested that miR-146a rs2910164 (C/G) locus was predicted to influence the risk of cancer. However, the relationship of miR-146a rs2910164 locus with colorectal cancer (CRC) susceptibility was controversial. We recruited 1003 CRC patients and 1303 controls, and performed a case-control study to clarify the correlation of miR-146a rs2910164 locus with CRC risk. Subsequently, a comprehensive meta-analysis was conducted to verify our findings. In the case-control study, we suggested that miR-146a rs2910164 variants did not alter CRC risk (CG vs. CC: adjusted P=0.465; GG vs. CC: adjusted P=0.436, CG/GG vs. CC: adjusted P=0.387 and GG vs. CC/CG: adjusted P=0.589), even in subgroup analysis. Next, we conducted a pooled-analysis to identify the correlation of miR-146a rs2910164 locus with CRC risk. In this pooled-analysis, 7947 CRC cases and 12,168 controls were included. We found that miR-146a rs2910164 polymorphism did not influence the risk of CRC (G vs. C: P=0.537; GG vs. CC: P=0.517, CG/GG vs. CC: P=0.520 and GG vs. CC/CG: P=0.167). Our findings suggest that miR-146a rs2910164 C/G polymorphism is not correlated with the susceptibility of CRC. In the future, more case-control studies are needed to confirm our results.

Methylenetetrahydrofolate reductase C677T (Ala>Val, rs1801133 C>T) polymorphism decreases the susceptibility of hepatocellular carcinoma: a meta-analysis involving 12,628 subjects.

C677T (Ala>Val, rs1801133 C>T), a non-synonymous variant of methylenetetrahydrofolate reductase (MTHFR) gene, has been found to be associated with an impair enzyme activity of MTHFR. The relationship of MTHFR rs1801133 with hepatocellular carcinoma (HCC) has been extensively investigated. However, the findings were conflicting. Recently, more investigations have been conducted on the relationship of MTHFR rs1801133 with HCC. To obtain a more precise assessment on the effect of this non-synonymous variant to the development of HCC, a pooled-analysis was performed. This meta-analysis consisted of 19 independent case-control studies. By using the odds ratio (OR) combined with 95% confidence interval (CI), the relationship of MTHFR rs1801133 with HCC risk was determined. A total of 19 independent case-control studies were included. Finally, 6,102 HCC cases and 6,526 controls were recruited to examine the relationship of MTHFR rs1801133 with HCC risk. In recessive model (TT vs. CC/CT), the findings reached statistical significance (OR, 0.90; 95%CI, 0.82-0.98; P = 0.016). Subgroup analysis also found an association between MTHFR rs1801133 polymorphism and the decreased risk of HCC in hepatitis/virus related patients (recessive model: OR, 0.85; 95%CI, 0.72-0.99; P = 0.035, and allele model: OR, 0.90; 95%CI, 0.81-0.99; P = 0.028). Subgroup analyses indicated that extreme heterogeneity existed in Asian population, larger sample size investigation, hospital-based study and normal/healthy control subgroups. The shape of Begger's seemed symmetrical. Egger's linear regression test also confirmed these evaluations. Sensitivity analyses suggested that our findings were stable. In summary, our results highlight that MTHFR rs1801133 polymorphism decreases HCC susceptibility. The relationship warrants a further assessment.

Association between methylenetetrahydrofolate reductase tagging polymorphisms and susceptibility of hepatocellular carcinoma: a case-control study.

Polymorphisms in one-carbon metabolism genes may influence the susceptibility to hepatocellular carcinoma (HCC). In the present study, we studied methylenetetrahydrofolate reductase (MTHFR) tagging polymorphisms in 584 HCC cases and 923 controls. Polymerase chain reaction was harnessed to detect MTHFR genotype. Overall, our results showed that genotype distribution of MTHFR rs4846048 and rs4845882 polymorphisms was not different between HCC patients and controls. MTHFR rs9651118 and rs1801133 loci were protective factors for HCC (rs9651118: CT vs. TT: adjusted odds ratio (OR) = 0.67, 95% confidence interval (CI): 0.49-0.90, P=0.008 and TC/CC vs. TT: adjusted OR = 0.70, 95% CI: 0.53-0.93, P=0.015; rs1801133: GA vs. GG: adjusted OR = 0.72, 95% CI: 0.54-0.97, P=0.031, AA/GA vs. GG: adjusted OR = 0.76, 95% CI: 0.57-0.99, P=0.045). However, MTHFR rs3753584 locus was a candidate for susceptibility to HCC (CT vs. TT: adjusted OR = 1.67, 95% CI: 1.20-2.32, P=0.003 and TC/CC vs. TT: adjusted OR = 1.59, 95% CI: 1.15-2.20, P=0.005). Results of haplotype analysis suggested that MTHFR Grs1801133Trs3753584Grs4845882Ars4846048Trs9651118 was associated with the risk of HCC (OR = 1.55, 95% CI: 1.16-2.07, P=0.003). The power of our study also confirmed these associations (the value of power >0.80). In summary, our findings suggested that MTHFR rs3753584, rs9651118 and rs1801133 polymorphisms may affect the risk of HCC in Chinese Han population. In future, our findings should be further validated in additional case-control studies.

Relationship of PPARG, PPARGC1A, and PPARGC1B polymorphisms with susceptibility to hepatocellular carcinoma in an eastern Chinese Han population.

BACKGROUND: PPARG, PPARGC1A, and PPARGC1B polymorphisms may be implicated in the development of cancer. PARTICIPANTS AND METHODS: In this study, we selected PPARG rs1801282 C>G and rs3856806 C>T, PPARGC1A rs2970847 C>T, and PPARGC1B rs7732671 G>C and rs17572019 G>A single-nucleotide polymorphisms to explore the relationship between these polymorphisms and hepatocellular carcinoma (HCC) risk. A total of 584 HCC patients and 923 controls were enrolled. RESULTS: We found that PPARG rs1801282 C>G polymorphism was correlated with a decreased susceptibility of HCC (CG vs CC, adjusted OR 0.47, 95% CI 0.27-0.82, P=0.007; CG/GG vs CC, adjusted OR 0.52, 95% CI 0.31-0.88, P=0.015). However, PPARG rs3856806 C>T polymorphism was a risk factor for HCC (TT vs CC, adjusted OR 2.33, 95% CI 1.25-4.36, P=0.008; TT vs CT/CC, adjusted OR 2.26, 95% CI 1.22-4.17, P=0.010). In a subgroup analysis by chronic hepatitis B virus (HBV)-infection status, age, sex, alcohol use, and smoking status, a significant association between PPARG rs1801282 C>G polymorphism and a decreased risk of HCC in male, ≥53 years, never-smoking, never-drinking, and nonchronic HBV-infection-status subgroups was found. However, we found PPARG rs3856806 C>T polymorphism increased the risk of HCC in never-smoking, never-drinking, and nonchronic HBV-infection-status subgroups. Haplotype-comparison analysis indicated that Crs1801282Trs3856806Crs2970847Grs7732671Grs17572019, Crs1801282Trs3856806Trs2970847Grs7732671Grs17572019, and Crs1801282Crs3856806Crs2970847Crs7732671Ars17572019 haplotypes increased the risk of HCC. PPARG Crs1801282Trs3856806 and Grs1801282Crs3856806 haplotypes also influenced the risk of HCC. CONCLUSION: In conclusion, our findings suggest PPARG polymorphisms may influence the susceptibility of HCC. The PPARG, PPARGC1A, and PPARGC1B haplotypes might be associated with HCC risk.

Investigation of LEP and LEPR polymorphisms with the risk of hepatocellular carcinoma: a case-control study in Eastern Chinese Han population.

BACKGROUND: Leptin (LEP) and LEP receptor (LEPR) polymorphisms may be associated with the development of cancer. METHODS: In this study, we selected five functional LEP and LEPR single-nucleotide polymorphisms (SNPs) and conducted a case-control study to determine the relationship of LEP and LEPR polymorphisms with hepatocellular carcinoma (HCC) risk in Eastern Chinese Han population. There were 584 HCC cases and 923 cancer-free controls included in our study. HCC patients and controls were fully matched by age and sex. SNPscan™ genotyping method was used to analyze the genotyping of LEP rs2167270 G>A, rs7799039 A>G, LEPR rs6588147 G>A, rs1137100 G>A, and rs1137101 G>A SNPs. RESULTS: We found that LEP rs7799039 A>G and rs2167270 G>A polymorphisms were associated with the susceptibility of HCC in this population (LEP rs7799039 A>G: GG vs AA: adjusted odds ratio [OR]=2.03, 95% CI, 1.22-3.38, P=0.006 and GG vs AA/AG: adjusted OR=1.97, 95% CI, 1.20-3.22, P=0.007; rs2167270 G>A: AA vs GG: adjusted OR=2.03, 95% CI, 1.10-3.75, P=0.024 and AA vs GG/GA: adjusted OR=2.01, 95% CI, 1.10-3.68, P=0.023). However, LEPR rs6588147 G>A polymorphism decreased the risk of HCC (GA vs GG: adjusted OR=0.62, 95% CI, 0.45-0.86, P=0.005 and AA/GA vs GG: adjusted OR=0.64, 95% CI, 0.47-0.88, P=0.007). CONCLUSION: This case-control study highlights that LEP rs7799039 A>G and rs2167270 G>A polymorphisms increase the susceptibility to HCC; however, LEPR rs6588147 G>A polymorphism may be a protective factor for HCC in Eastern Chinese Han population.

Association between ICOS polymorphisms and risk of colorectal cancer: a case-control study involving 2,606 subjects.

Functional variants in inducible T cell costimulator (ICOS) gene are predicted to be associated with the susceptibility of colorectal cancer (CRC). In this study, we enrolled 2,606 participants (involving 1,003 CRC cases and 1,303 healthy controls) and conducted a case-control study to explore the potential relationship of ICOS rs4404254 T>C and rs10932029 T>C polymorphisms with the risk of CRC. A custom-by-design 48-Plex SNPscan Kit was used to obtain the genotypes of ICOS rs4404254 T>C and rs10932029 T>C variants. We found that ICOS rs10932029 T>C polymorphism was associated with risk of CRC in several subgroups (female subgroup: CC vs. TT: adjusted OR = 6.49, 95% CI 1.36-30.90, P = 0.019 and CC vs. CT/TT: adjusted OR = 6.38, 95% CI 1.34-30.32, P = 0.020; < 61 years subgroup: CC vs. TT: adjusted OR = 4.23, 95% CI 1.10-16.24, P = 0.036 and CC vs. CT/TT: adjusted OR = 4.20, 95% CI 1.10-16.09, P = 0.036; never smoking subgroup: CC vs. TT: adjusted OR = 2.82, 95% CI 1.04-7.64, P = 0.041 and CC vs. CT/TT: adjusted OR = 2.83, 95% CI 1.05-7.66, P = 0.041 and BMI ≥ 24 subgroup: CC vs. TT: adjusted OR = 6.81, 95% CI 1.39-33.30, P = 0.018 and CC vs. CT/TT: adjusted OR = 6.79, 95% CI 1.39-33.11, P = 0.018). In addition, we found that ICOS rs4404254 T>C polymorphism was associated with the susceptibility of CRC in never smoking subgroup (CC/TC vs. TT: adjusted OR = 1.23, 95% CI 1.01-1.51, P = 0.045). In summary, our findings suggest that ICOS rs10932029 T>C and ICOS rs4404254 T>C polymorphisms may be associated with the risk of CRC. In the future, a fine-mapping study with a functional evaluation is needed to explore the relationship between ICOS polymorphisms and the risk of CRC.

Association of PPARG rs 1801282 C>G polymorphism with risk of colorectal cancer: from a case-control study to a meta-analysis.

The functional single nucleotide polymorphisms in peroxisome proliferator-activated receptor gamma (PPARG) gene were predicted to be correlated with the susceptibility of colorectal cancer (CRC). The aim of the present study was to explore the relationship between PPARG rs1801282 C>G polymorphism and the risk of CRC. First, we conducted a case-control study with 387 CRC cases and 1,536 controls. We used the SNPscan method to determine the genotypes of PPARG rs1801282 C>G polymorphism. We found PPARG rs1801282 C>G polymorphism had a tendency of decreased risk to CRC risk (CG vs. CC: adjusted OR, 0.67, 95% CI = 0.43-1.04 for CG vs. CC, P = 0.073; GG vs. CC: adjusted OR, 0.68; 95% CI, 0.44-1.05; P = 0.078). The stratified analysis revealed PPARG rs1801282 C>G polymorphism also had a tendency of decreased risk to colon cancer (CG vs. CC: adjusted OR = 0.54, 95% CI = 0.27-1.08, P = 0.083). The results of subsequent meta-analysis suggested that PPARG rs1801282 C>G polymorphism might be a protective factor for CRC, especially in Asians, colon cancer and rectum cancer subgroups. In conclusion, our study indicates that PPARG rs1801282 C>G polymorphism might decrease the risk of overall CRC. Larger sample size and well-designed case-control studies are needed to confirm the potential association.

Epidermal growth factor induces HCCR expression via PI3K/Akt/mTOR signaling in PANC-1 pancreatic cancer cells.

BACKGROUND: Human cervical cancer oncoprotein 1 (HCCR-1), reported as a negative regulator of p53, is over-expressed in a variety of human cancers. However, it is yet unknown whether HCCR-1 plays any role in pancreatic cancer development. The aim of this study was to investigate the effect of epidermal growth factor on the expression of HCCR in pancreatic cancer cells, and to explore if PI3K/Akt/mTOR signaling pathway mediated this expression. METHODS: A polyclonal antibody against HCCR protein was raised by immunizing Balb/c mice with the purified recombinant protein pMBPc-HCCR. Tissue samples were constructed on a tissue chip, and the expression of HCCR was investigated by immunohistochemistry assay and Western blotting. Pancreatic cell line, PANC-1 cells were stably transfected with plasmids containing sense-HCCR-1 fragment and HCCR siRNA fragment. MTT and transwell assay were used to investigate the proliferation and invasion of stable tansfectants. The specific inhibitor of PI3K and mTOR was used to see if PI3K/mTOR signal transduction was involved in the induction of HCCR gene expression. A Luciferase assay was used to see if Akt can enhance the HCCR promoter activity. RESULTS: HCCR was up-regulated in pancreatic tumor tissues (mean Allred score 4.51+/-1.549 vs. 2.87+/-2.193, P<0.01), especially with high expression in poorly differentiated pancreatic cancer. The growth of cells decreased in HCCR-1 siRNA transfected cells compared with vector transfectants. The number of invasion cells was significantly lower in HCCR-1 siRNA transfected cells (24.4+/-9.9) than that in vector transfectants (49.1+/-15.4). Treatment of PANC-1 cells with epidermal growth factor increased HCCR protein level in a dose- and time-dependent manner. However, application of LY294002 and rapamycin caused a dramatic reduction of epidermal growth factor-induced HCCR expression. Over-expression of exogenous constitutively active Akt increased the HCCR promoter activity; in contrast, dominant negative Akt decreased the promoter activity. CONCLUSIONS: EGF-induced HCCR-1 over-expression is mediated by PI3K/AKT/mTOR signaling which plays a pivotal role in pancreatic tumor progression, suggesting that HCCR-1 could be a potential target for cancer therapeutics.