Activation of the PI3K/AKT pathway could reflect phosphorylation levels of AKT proteins and after phosphorylation, it could be further activated a variety of downstream proteins, such as p21, p27 and caspase-3, which could regulate the state of tumor cells

Activation of the PI3K/AKT pathway could reflect phosphorylation levels of AKT proteins and after phosphorylation, it could be further activated a variety of downstream proteins, such as p21, p27 and caspase-3, which could regulate the state of tumor cells. and the xenograft tumor tissue samples were analyzed for the expression of PCNA and Ki-67 by immunohistochemistry and the cell morphology was evaluated by hematoxylin and eosin (H&E). Results revealed that hnRNP A2/B1 was successfully silenced in HeLa and CaSki cells. hnRNP A2/B1 knock-down significantly induced the suppression of proliferation, migration, invasion and also enhancement of apoptosis and reduced the IC50 of lobaplatin and irinotecan. The expression of p21, p27 and cleaved caspase-3 in shRNA group were significantly upregulated and the expression of p-AKT was reduced both and and and the brown particles were labeled as positive areas. In addition, H&E staining was used to observe the morphological structure in tumor tissues. The results suggested that the positive expression of PCNA (P<0.05) and Ki-67 (P<0.01) were significantly lower in hnRNP A2/B1 knockdown tumor group compared to the other group (Fig. 9C and Table IV). As shown in Fig. 9D, the characteristics of xenograft tissues conformed to tumor cells and were as follows: Acidophil hepatocytes with both nuclear and cytoplasmic enlargement, nuclear pleomorphism and hyperchromasia, and frequent multinucleation. In order to further demonstrate the relationship between the PI3K/AKT signaling pathway and hnRNP A2/B1 in nude mouse xenograft tissues, western blotting was used for clarification. The xenograft tumor of hnRNP A2/B1-shRNA group could suppress the expression of p-AKT protein, upregulating cleaved caspase-3, p21 and p27 (Fig. 9E). The results indicated that it was consistent with the earlier apoptotic and cycle results from the protein level of xenograft tumor tissues. Open in a separate window Figure 9. hnRNP A2/B1 knockdown inhibits the growth of cervical NU 1025 cancer HeLa cells and at hnRNP A2/B1 downregulation group and the result suggested that the hnRNP A2/B1 affected NU 1025 cell cycle by regulated p21 and p27 in cervical cancer. Previous studies showed that hnRNP A2/B1 can upregulate the proportion of anti-apoptosis factors and proteins in cells to promote the malignant growth of tumors (41), NU 1025 our study also confirmed this argument. Caspase-3 may be involved in cell apoptosis (42), our results indicated that silencing hnRNP A2/B1 enhanced apoptosis in cervical cancer via activation of caspase-3. Aberrant activation of the PI3K/AKT pathway is widespread in malignant tumors and is an important pathway to mediate cell cycle, and apoptosis (43,44). Licochalcone A induced autophagy by inactivation of PI3K/AKT/mTOR pathway in cervical cancer cells (45). Activation of the PI3K/AKT pathway could reflect phosphorylation levels of AKT proteins and after phosphorylation, it could be further activated a variety of downstream proteins, such as p21, p27 and caspase-3, which could regulate the state of tumor cells. Our results demonstrated that the expression of p-AKT was reduced in hnRNP A2/B1 knockdown group both and and hnRNP A2/B1 was related to PI3K/AKT pathway in promotion of cervical cancer. Previous studies have reported that hnRNP A2/B1 regulates the self-renewal, cell cycle and pluripotency in human embryonic stem cells is related to PI3K/AKT pathway (46) and this was similar to our results. In conclusion, our findings demonstrate that inhibiting hnRNP A2/B1 expression in cervical cancer can induce NSHC apoptosis and cell cycle arrest and enhance the chemotherapy sensitivity of cervical cancer cells to NU 1025 lobaplatin and irinotecan. Analysis of cervical cancer cell lines HeLa and CaSki cells shows that hnRNP A2/B1 knockdown can reduce the ability of cell proliferation, invation and migration, indicating that hnRNP A2/B1 may be one of the central regulators for cervical cancer. The activation of PI3K/AKT pathway is one of the important mechanisms for hnRNP A2/B1 to facilitate the development of cervical cancer..