TGFβ3-mediated induction of Periostin facilitates head and neck cancer growth and is associated with metastasis

The matrix-specific protein periostin (POSTN) is up-regulated in human cancers and associated with cancer growth, metastasis and angiogenesis. Although the stroma of cancer tissues is the main source of POSTN, it is still unclear how POSTN plays a role to facilitate the interplay between cancer cells and cancer-associated fibroblasts (CAFs) in head and neck cancer (HNC), thereby promoting tumorigenesis via modifying the tumor microenvironment. Herein, we have performed studies to investigate POSTN and its role in HNC microenvironment. Our results indicated that POSTN was significantly up-regulated in HNCs, especially in the tissues with lymph node metastasis. Moreover, POSTN was highly enriched in the stroma of cancer tissues and produced mainly by CAFs. More importantly, we have pinpointed TGF-β3 as the major upstream molecular that triggers the induction of POSTN in CAFs. As such, during the interaction between fibroblasts and cancer cells, the increased stromal POSTN induced by TGF-β3 directly accelerated the growth, migration and invasion of cancer cells. Hence, our study has provided a novel modulative role for POSTN on HNC progression and further reveals POSTN as an effective biomarker to predict metastasis as well as a potential cancer therapeutic target

Human Papillomavirus Type 18 E6 and E7 Genes Integrate into Human Hepatoma Derived Cell Line Hep G2

Background and Objectives: Human papillomaviruses have been linked causally to some human cancers such as cervical carcinoma, but there is very little research addressing the effect of HPV infection on human liver cells. We chose the human hepatoma derived cell line Hep G2 to investigate whether HPV gene integration took place in liver cells as well. Methods: We applied PCR to detect the possible integration of HPV genes in Hep G2 cells. We also investigated the expression of the integrated E6 and E7 genes by using RT-PCR and Western blotting. Then, we silenced E6 and E7 expression and checked the cell proliferation and apoptosis in Hep G2 cells. Furthermore, we analyzed the potential genes involved in cell cycle and apoptosis regulatory pathways. Finally, we used in situ hybridization to detect HPV 16/18 in hepatocellular carcinoma samples. Results: Hep G2 cell line contains integrated HPV 18 DNA, leading to the expression of the E6 and E7 oncogenic proteins. Knockdown of the E7 and E6 genes expression reduced cell proliferation, caused the cell cycle arrest at the S phase, and increased apoptosis. The human cell cycle and apoptosis real-time PCR arrays analysis demonstrated E6 and E7-mediated regulation of some genes such as Cyclin H, UBA1, E2F4, p53, p107, FASLG, NOL3 and CASP14. HPV16/18 was found in only 9% (9/100) of patients with hepatocellular carcinoma. Conclusion: Our investigations showed that HPV 18 E6 and E7 genes can be integrated into the Hep G2, and we observed a low prevalence of HPV 16/18 in hepatocellular carcinoma samples. However, the precise risk of HPV as causative agent of hepatocellular carcinoma needs further study

Preparation and application research of hybrid alkali-resistant glass fiber reinforced cement grout

Cement grouting reinforcement is a widely used method for controlling water inrush in tunnels. However, traditional cement grout has certain limitations such as low strength and susceptibility to erosion, which can result in inadequate grouting performance in water-rich tunnels. To address this issue, this study introduces a novel type of grout called hybrid alkali-resistant glass fiber reinforced cement grout (HARGFC). In order to evaluate its effectiveness, different ratios of HARGFC were tested in terms of slurry fluidity, splitting tensile strength, compressive strength, and flexural strength through laboratory experiments. Additionally, microscopic analysis of the samples was carried out using scanning electron microscopy (SEM) in conjunction with the mechanical tests. Finally, the application of HARGFC was demonstrated in a water-blocking grouting project conducted in the Qiyue Mountain Tunnel on the Liwan Expressway. The results demonstrated that this material possesses outstanding physical and mechanical properties, as well as alkali resistance, which greatly enhances grouting performance in water-flowing tunnels. Adding a total fiber content of 1%, that is, 80% of Cem-FIL70 (9 mm) and 20% of HD (6 mm), yielded the optimal ratio for HARGFC. Compared to traditional cement materials, the splitting tensile strength, flexural strength, and compressive strength of the new material increased by 27.46%, 20.64%, and 7.37%, respectively. Building upon various combinations of grouting methods, the optimal HARGFC was applied to the grouting sealing and reinforcement of the Qiyue Mountain Tunnel, effectively controlling water inflow. These research findings provide valuable data and theoretical support for addressing sudden water inrush in tunnels and grouting construction

siRNA targeting HPV 18 E7 sites and sequences.

<p>siRNA targeting HPV 18 E7 sites and sequences.</p

Inhibition of HPV 18 E7 gene inhibited cell growth in Hep G2.

<p>(A) Hep G2 cells transfected with E7-siRNA or control siRNA were evaluated in EdU assays at 0 h, 12 h, 24 h, 36 h, 48 h, 60 h and 72 h after transfection. After transfection of Hep G2 cells with E7-siRNA, a time-dependent reduction of cell proliferation was observed at 48 h, 60 h and 72 h. (B) At 48 h, 60 h and 72 h following E7-siRNA transfection, the percents of S-phase cells were 9.39%±3.55%, 17.29%±5.85% and 30.87%±4.26% compared with 18.74%±6.66%, 24.03%±5.35% and 41.97%±8.73% in NC-siRNA transfected cells (p<0.05).</p

mRNA expression of E7-related genes in Hep G2 cells after the silencing of HPV 18 E7 through RT² Profiler™ Human Cell Cycle PCR Array.

<p>In 84 E7-related genes, 19 genes mRNA transcript were altered markedly compared with the negative control. Standards of eligibility: folds up- or down-regulation >2.0 and <i>p</i><0.05; n = 3.</p

Transfection with E7-siRNA induced apoptosis.

<p>(A, B, C and D) The percent of apoptotic Hep G2 cells was measured using the Annexin V assay at 24 h, 48 h and 72 h after transfection, and then the stained cells were analyzed through flow cytometry. (E) The results showed that 7.26%±0.29%, 22.03%±0.23% and 19.20%±0.78% in siRNA E7 transfected Hep G2 cells after 24 h, 48 h and 72 h underwent total apoptosis compared with only 5.25%±0.76% in NC siRNA E7 transfected cells (p<0.05).</p

mRNA expression of E7-related genes in Hep G2 cell after the silencing of HPV 18 E7 in RT² Profiler™ Human Apoptosis PCR Array.

<p>In 84 E7-related genes, 21 genes mRNA transcript were altered markedly compared with the negative control. Standards of eligibility: folds up- or down-regulation >2.0 and <i>p</i><0.05; n = 3.</p