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Gene & Protein in Disease m1A-mediated ESCCAL-1 promotes ESCA stemness

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D E F

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Figure 4. ESCCAL-1 is regulated by ALKBH3 in an m A-dependent manner. (A) The expression levels of ALKBH3 and ESCCAL-1 in 13 normal tissues
and 182 ESCA tissues were analyzed by the GEPIA database. *P < 0.05. (B) The GEPIA database was used to investigate the expression correlation between
ALKBH3 and ESCCAL-1 in 182 ESCA samples. (C) The mRNA levels of ALKBH3 in Het-1A, TE1, KYSE70, and EC1 cells were tested by qRT-PCR, n = 4.
***P < 0.001. (D and E) The effect of ALKBH3 silencing on the expression of ALKBH3 and ESCCAL-1 in ESCA cells was evaluated by qRT-PCR, n = 3.
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*P < 0.05, **P < 0.01, ***P < 0.001. (F) The putative presence of m A-modified regions on ESCCAL-1 transcripts. (G) MeRIP-PCR assay was used to detect
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m A modification on ESCCAL-1 transcripts, n = 3. **P < 0.01. (H) MeRIP-PCR combined with agarose gel electrophoresis was used to evaluate the effect
of ALKBH3 knockdown on m A modification of ESCCAL-1 transcript, n = 2.
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ESCCAL-1: Esophageal squamous cell carcinoma associated long non-coding RNA 1; ESCA: Esophageal cancer; m A: N -methyladenosine.
Stem cell-like property refers to the self-renewing contrast, overexpression of ESCCAL-1 caused the opposite
ability of malignant tumor cells to maintain continuous biological effects. Although we have not clarified the
proliferation and metastasis [22,23] . The stemness nature of exact mechanisms by which ESCCAL-1 is involved in the
tumors allows them to adapt to extreme growth conditions maintenance of ESCA stemness, our results preliminarily
and to combat multiple risk factors [22,23] . Therefore, reducing suggest that ESCCAL-1 may exert its function by regulating
tumor stemness is critical in eliminating tumor therapeutic the expression of CD44, KLF4, and other stemness
resistance [24,25] . In the previous studies, ESCCAL-1 has the markers. This study provides new insights into targeting the
carcinogenic effect of promoting ESCA proliferation and ESCCAL-1/stemness axis for ESCA treatment.
metastasis [9,10] , two basic intrinsic properties of malignant In addition to the high expression of ESCCAL-1 in
tumors, and depend on a high degree of self-renewal ESCA, we also found that ESCCAL-1 is closely related to
ability. Therefore, ESCCAL-1 might be associated with tumor stage and patient survival, further supporting the
the stemness of ESCA. Through a series of functional and view that ESCCAL-1 is a potential biomarker of ESCA.
molecular expression experiments, we demonstrated that In addition, we noted that this lncRNA had a special
the knockdown of ESCCAL-1 significantly weakened the relationship with the gender and race of ESCA patients. Its
stemness of ESCA cells and reduced the mRNA and protein expression level in male tumor samples was higher than
levels of two stemness-related genes, CD44 and KLF4. In that in female tumor samples, and its expression level in

Volume 2 Issue 1 (2023) 7 https://doi.org/10.36922/gpd.305