SP1/RNASEH2A accelerates the development of hepatocellular carcinoma by regulating EMT

Background: The expression level of Ribonuclease H2, subunit A (RNASEH2A) in hepatocellular carcinoma (HCC) has been reported, but the function of RNASEH2A on HCC cells development and the related molecular mechanisms remain unclear. Herein, we intend to explore the upstream regulator of RNASEH2A and its role in the HCC progression. Methods: GEPIA website was employed to determine the level of RNASEH2A in HCC tissues and get a survival analysis. After reducing RNASEH2A expression by RNA interference, cell counting kit-8, colony formation, Western blot, Transwell and wound healing assays were performed to estimate the malignant properties of HCC cells. The transcriptional factor of RNASEH2A was predicted by UCSC and JASPAR database and confirmed by dual luciferase assay and Ch-IP assay. The expression level of EMT pathway related molecules was determined by western blotting. Results: An increased expression of RNASEH2A was presented in HCC and predicted worse prognosis of HCC patients. Functionally, the results demonstrated that depletion of RNASEH2A suppressed HCC cell proliferation, cell cycle, migration and invasion. Moreover, we illustrated that SP1 targeted to the promoter of RNASEH2A and modulated its expression in HCC cell lines. RNASEH2A knockdown counteracted the function of SP1 overexpression in modulating HCC cell growth, cell cycle, and mobility. Then, our data showed that the SP1/RNASEH2A axis affected the malignant behaviors of HCC cells by regulating EMT process. Conclusions: In summary, these results demonstrated that RNASEH2A promoted HCC cells development through regulating EMT process and was transcriptionally modulated by SP1

Electron Beam Welding and Post Heat Treatment of a New Near-Beta High-Strength Ti-4Al-5Mo-5V-5Cr-1Nb Alloy

Ti-4Al-5Mo-5V-5Cr-1Nb (wt.%) is a new type of high-strength (~1300 MPa) titanium (Ti) alloy developed for aerospace applications. Until now, the research on its welding and subsequent heat treatment is barren. Herein, we employed electron beam welding (EBW) to a solutionized Ti-4Al-5Mo-5V-5Cr-1Nb with a phase constituent of α + β and investigated its microstructure and mechanical properties in both as-welded (AW) and post-weld aging treated (PWAT) conditions. Results showed that due to the thermal input of the welding process, the α phase in the original microstructure of base material (BM) transformed into the β phase in the fusion zone (FZ). Similar microstructural evolution was observed for the heat-affected zone (HAZ) near the FZ (Near-HAZ), whereas the HAZ far away from FZ (Far-HAZ) contained a small amount of round α phase (ghost α) due to its slower cooling rate. Such a microstructural change resulted in poor tensile strength (~780 Mpa) for the as-welded joint. After PWAT, a large number of acicular α precipitated in the FZ and HAZ and its size (S) in different zones followed the order of SFar-HAZ FZ ≈ SNear-HAZ BM. The presence of αs precipitates remedied the tensile strength of the weld joint almost to the same as that of the BM. The present findings established the foundation of the application of this high-strength Ti alloy

KRN4 Controls Quantitative Variation in Maize Kernel Row Number.

Kernel row number (KRN) is an important component of yield during the domestication and improvement of maize and controlled by quantitative trait loci (QTL). Here, we fine-mapped a major KRN QTL, KRN4, which can enhance grain productivity by increasing KRN per ear. We found that a ~3-Kb intergenic region about 60 Kb downstream from the SBP-box gene Unbranched3 (UB3) was responsible for quantitative variation in KRN by regulating the level of UB3 expression. Within the 3-Kb region, the 1.2-Kb Presence-Absence variant was found to be strongly associated with quantitative variation in KRN in diverse maize inbred lines, and our results suggest that this 1.2-Kb transposon-containing insertion is likely responsible for increased KRN. A previously identified A/G SNP (S35, also known as Ser220Asn) in UB3 was also found to be significantly associated with KRN in our association-mapping panel. Although no visible genetic effect of S35 alone could be detected in our linkage mapping population, it was found to genetically interact with the 1.2-Kb PAV to modulate KRN. The KRN4 was under strong selection during maize domestication and the favorable allele for the 1.2-Kb PAV and S35 has been significantly enriched in modern maize improvement process. The favorable haplotype (Hap1) of 1.2-Kb-PAV-S35 was selected during temperate maize improvement, but is still rare in tropical and subtropical maize germplasm. The dissection of the KRN4 locus improves our understanding of the genetic basis of quantitative variation in complex traits in maize

The four polymorphisms in <i>KRN4</i> and <i>UB3</i> associated with KRN under the MLM K + Q model.

<p>^a The alleles represent ‘desirable allele/undesirable allele’.</p><p>The four polymorphisms in <i>KRN4</i> and <i>UB3</i> associated with KRN under the MLM K + Q model.</p

Expression analysis and phenotypic characterization of <i>UB3-mum4</i> and <i>UB2-mum3</i> mutants.

<p>A) The insertion site of <i>Mutator</i> in <i>UB3-mum4</i> and <i>UB2-mum3</i> mutants. B and C) Expression level of <i>UB3</i> (B) and <i>UB2</i> (C) in mutant and wild type. ** P < 0.01, * P < 0.05. D) KRN performance single and double mutants of <i>UB3-mum4</i>/<i>UB2-mum3</i> and wild type; WH: Wuhan; SY: Sanya. E) Tassel and ear of wild type and double mutant of <i>UB3-mum4</i>/<i>UB2-mum3</i>. Detailed information regarding phenotypes is presented in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005670#pgen.1005670.s009" target="_blank">S3</a>–<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005670#pgen.1005670.s012" target="_blank">S6</a> Tables.</p

Fine mapping of <i>KRN4</i>.

<p>A) The graphical genotypes of homozygous recombinants (HR). The white boxes in the graphical genotype represent the genomic segments from H21, the black boxes represent genomic segments from H21^NX531. A progeny test was conducted to examine whether the KRN of HRs were significantly higher than that of H21. N: the total number of HR phenotyped in four environments (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005670#pgen.1005670.s014" target="_blank">S1 Dataset</a>). <i>P</i>-value: Student’s t-test of the difference in KRN between HRs and H21. The axis represents the physical map of <i>KRN4</i>, and the blue box represents genes within <i>KRN4</i>. B) Nucleotide sequence differences in the <i>KRN4</i> region between H21 and H21^NX531. The orange solid boxes represent the locations of transposable elements in <i>KRN4</i>.The shadowed regions represent homologous <i>KRN4</i> sequence between H21 and H21^NX531 (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005670#pgen.1005670.s015" target="_blank">S2 Dataset</a>).</p

Pleiotropic effects of <i>KRN4</i>.

<p>^aN, sample size, H21/H21^NX531</p><p>Pleiotropic effects of <i>KRN4</i>.</p

Analysis of <i>UB3</i> and GRMZM2G001541 expression.

<p>A) Expression patterns of <i>UB3</i> and GRMZM2G001541 (<i>GRM541</i>) in immature ears of H21 and H21^NX531, **: <i>P</i>-value < 0.01. B) Analysis of <i>UB3</i> and GRMZM2G001541 (<i>GRM541</i>) expression in recombinant lines in immature 2-mm ear. The white boxes in the graphical genotype represent the genomic segment from H21, and the black boxes represent the genomic segment from H21^NX531. C) Expression of <i>UB3</i> in immature 2-mm ear in 38 diverse inbred lines. H represents lines with the H21^NX531genotype at the <i>KRN4</i> locus (N = 12), and L represents lines with the H21genotype at the <i>KRN4</i> locus (N = 26).</p

KRN and frequencies of haplotypes between 1.2-Kb PAV and S35.

<p>^a The sample size for all maize is 428</p><p>^b TST: <u>T</u>ropical and <u>S</u>ub<u>T</u>ropical maize germplasm, sample size: 234</p><p>^c TEMP: <u>Temp</u>erate maize germplasm, sample size: 194.</p><p>KRN and frequencies of haplotypes between 1.2-Kb PAV and S35.</p

The evidence of significant selection in <i>KRN4</i> during maize domestication and improvement.

<p>A) Nucleotide diversities (π) within ~3 Kb of <i>KRN4</i> for maize landrace (red line) and teosinte (blue line). The green shades represent the 1.2-Kb PAV region. **: <i>P</i>-value < 0.01. B) Allele frequencies of <i>1</i>.<i>2-Kb Presence</i> of 1.2-Kb PAV in teosinte, maize landrace and inbred lines. C) Allele frequencies of <i>A</i> of S35 in teoisnte, maize landrace and inbred lines. D) A putative evolutionary pattern of 1.2-Kb PAV and S35 during maize domestication and improvement. The circles in colors represent the four haplotypes between 1.2-Kb PAV and S35. The genotypes of haplotypes in 1.2-Kb PAV and S35 are showed in the circles. The number inside the circle is the frequency of the haplotype. TST maize: <u>T</u>ropical and <u>S</u>ub<u>T</u>ropical maize germplasms. All of the steps marked as “Selection” are of significantly frequency change, <i>P</i>-value < 0.001 (χ² test based on the frequencies).</p