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Expression and clinical importance of a newly discovered alternative splice variant of the gene for acrosin binding protein found in human brain tumors

Baolong Zheng
Baolong Zheng
   | 31 dic 2020
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Article Category: Original article
Pubblicato online: 31 dic 2020
Pagine: 243 - 252
DOI: https://doi.org/10.1515/abm-2020-0033
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© 2020 Baolong Zheng, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Nucleotide and predicted amino acid sequences for the variant of the gene for acrosomal binding protein, ACRBP-V5a. The nucleotide sequence is shown above the predicted amino acid sequence. Numbers on both sides correspond to nucleotides (roman) and amino acid residues (bold italic). The exon 5a is underlined. Two potential N-glycosylation sites are shaded (based on the NetNGlyc1.0 Server).
Nucleotide and predicted amino acid sequences for the variant of the gene for acrosomal binding protein, ACRBP-V5a. The nucleotide sequence is shown above the predicted amino acid sequence. Numbers on both sides correspond to nucleotides (roman) and amino acid residues (bold italic). The exon 5a is underlined. Two potential N-glycosylation sites are shaded (based on the NetNGlyc1.0 Server).

Figure 2

Expression of the variant of the gene for acrosomal binding protein, ACRBP-V5a and the gene for human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in tissues and cell lines. Left of the DNA ladder (lane M, catalog Nos. MD101-01 or MD104-01; Vazyme Biotech) are the respective numbers of base pairs. Lane N, reaction mixture with H2O alone. (A) Primers located around the human ACRBP-V5a cDNA region are encoded by exons 4–5a and bands of 600 bp represent the splice variant. Lane 1 cDNA of SHG-44 (glioma cell line), lanes 2 and 3 hep G2 and 3 SMMC7721 (both hepatocarcinoma cell lines), respectively. (B) Lanes S1–9 represent tumor tissues of 9 patients and cloned 496 bp of GAPDH cDNA. (C) Positive control testis sample (lane P) derived from SMMC7721 cDNA.
Expression of the variant of the gene for acrosomal binding protein, ACRBP-V5a and the gene for human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in tissues and cell lines. Left of the DNA ladder (lane M, catalog Nos. MD101-01 or MD104-01; Vazyme Biotech) are the respective numbers of base pairs. Lane N, reaction mixture with H2O alone. (A) Primers located around the human ACRBP-V5a cDNA region are encoded by exons 4–5a and bands of 600 bp represent the splice variant. Lane 1 cDNA of SHG-44 (glioma cell line), lanes 2 and 3 hep G2 and 3 SMMC7721 (both hepatocarcinoma cell lines), respectively. (B) Lanes S1–9 represent tumor tissues of 9 patients and cloned 496 bp of GAPDH cDNA. (C) Positive control testis sample (lane P) derived from SMMC7721 cDNA.

Figure 3

Sequence of the TA cloning result for the variant of the gene for acrosomal binding protein, ACRBP-V5a.
Sequence of the TA cloning result for the variant of the gene for acrosomal binding protein, ACRBP-V5a.

Figure 4

Genomic structure of the gene for acrosomal binding protein, ACRBP. The exons are shown as boxes and the introns as lines. The dark box is exon 5a. Two double-headed arrows represent the cloning sequences of E5, E8, and V5a respectively.
Genomic structure of the gene for acrosomal binding protein, ACRBP. The exons are shown as boxes and the introns as lines. The dark box is exon 5a. Two double-headed arrows represent the cloning sequences of E5, E8, and V5a respectively.

Figure 5

The ratios of ACRBP to ACRBP-V5a in the 3 types of brain tumors. Data represent mean ± SE. A Mann–Whitney U test was used to determine the differences between the 3 different types. N.S.: not significant; **P < 0.01, ***P < 0.001.
The ratios of ACRBP to ACRBP-V5a in the 3 types of brain tumors. Data represent mean ± SE. A Mann–Whitney U test was used to determine the differences between the 3 different types. N.S.: not significant; **P < 0.01, ***P < 0.001.

Figure 6

Comparison of porcine (full-length) and human (V5a) amino acid sequences for acrosomal binding protein (ACRBP; Clustal Omega alignment program, https://www.ebi.ac.uk/Tools/msa/clustalo/). An * (asterisk) indicates positions that have a single, fully conserved residue. A: (colon) indicates conservation between groups (porcine and human) of strongly similar properties. A. (period) indicates conservation between groups of weakly similar properties.
Comparison of porcine (full-length) and human (V5a) amino acid sequences for acrosomal binding protein (ACRBP; Clustal Omega alignment program, https://www.ebi.ac.uk/Tools/msa/clustalo/). An * (asterisk) indicates positions that have a single, fully conserved residue. A: (colon) indicates conservation between groups (porcine and human) of strongly similar properties. A. (period) indicates conservation between groups of weakly similar properties.

Analysis of ACRBP-V5a expression and clinicopathological characteristics

Clinicopathological characteristicNo. of mRNA positive patient samples/total samples (%)†PRelative expression Mean ± SE‡P
Sex
  Male21/63 (33.3)0.2946.7 ± 14.00.81
  Female13/29 (44.8)41.7 ± 11.3
Age (years)
  <3420/46 (43.5)0.2050.7 ± 9.50.47
  ≥3414/46 (30.4)36.4 ± 19.1
KPS§
  <7015/34 (44.1)0.2857.6 ± 18.70.24
  ≥7019/58 (32.8)34.7 ± 8.5
Tumor grade‖
  II10/33 (30.3)0.3217.7 ± 3.80.01*
  IV24/59 (40.7)56.1 ± 12.8
Tumor type
  Astrocytoma10/33 (30.3)0.3017.7 ± 3.80.02*
  Glioblastoma10/30 (33.3)50.4 ± 26.1
  Medulloblastoma14/29 (48.3)60.2 ± 12.7
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