Application of Hsp27 in tolerance diagnosis and treatment for tongue cancer chemotherapy

Abstract

The invention discloses an application of Hsp27 in tolerance diagnosis and treatment for tongue cancer chemotherapy. The conditions that the Hsp27 protein level in a tongue cancer cell culture supernatant and the Hsp27 protein level in serum of a tongue cancer patient can be raised through chemotherapeutic drug treatment and expression of the Hsp27 in a tolerance cell for tongue cancer chemotherapy is higher than that in a sensitive cell are verified for the first time; and the chemosensitivity of a drug-resistant cell is improved by interfering with and lowering the expression of the Hsp27 and the chemotherapy tolerance of the tongue cancer cell is improved through overexpression of the Hsp27. New data is provided for research on a tolerance mechanism for tongue cancer chemotherapy, a new detection marker is provided for prediction of the tongue cancer chemosensitivity, and a theoretical basis is provided for development of a new treatment strategy based on the Hsp27 as a target.

Description

technical field

[0001] The invention belongs to the field of biotechnology or medicine, and more specifically, the invention relates to the application of Hsp27 in the diagnosis and treatment of tongue cancer chemotherapy resistance. Background technique

[0002] Heat shock proteins are a group of highly conserved proteins in evolution. In addition to high fever, ischemia, tissue damage, virus or bacterial infection, etc. are also produced, and they have been proved to play various important functions in many fields. Previous studies on heat shock protein 27 (Heat shock protein 27, Hsp27) mainly focused on the anti-apoptosis and anti-oxidative free radical effects of Hsp27 (Small heat shock proteins and protection against injury. Ann N Y AcadSci. 199930; 874 : 66-8.Review); At present, there is no understanding and in-depth research on the application of Hsp27 in the diagnosis and treatment of tongue cancer chemotherapy resistance. Contents of the invention

[0003] In vi...

Examples

Embodiment 1

[0020] Example 1: Chemotherapy can induce the increase of Hsp27 in tongue cancer cells and serum of patients

[0021] The results showed that the level of Hsp27 protein in drug-resistant cells Tca8113 / PYM was significantly higher than that in normal oral mucosa cells Hacat and Tca8113, SCC-25, SCC-15, SCC-9 and CAL-27, which were relatively sensitive to chemotherapy ( Figure 1A ), and the Hsp27 protein level in the drug-resistant cell culture medium was also significantly higher than that of Hacat, Tca8113, SCC-25, SCC-15, SCC-9 and CAL-27 cell culture medium ( Figure 1A ); we treated the above cells with PYM (80mg / L) and cDDP (5mg / L) respectively, and found that chemotherapy could induce Hsp27 protein levels in each cell and in the cell culture medium to varying degrees ( Figure 1A ); In addition, we detected the protein level of Hsp27 in the serum of 30 cases of normal healthy people and 50 cases of tongue cancer patients. Serum of patients with tongue cancer found that ...

Embodiment 2

[0022] Example 2: In vitro experiment of Hsp27 increasing chemotherapy resistance of tongue cancer cells

[0023] In order to observe the role of Hsp27 in chemotherapy resistance of tongue cancer, we silenced the expression of Hsp27 in Tca8113 / PYM cells by shRNA technology, and the interference effects of sh-1# and sh-2# were the most obvious ( Figure 2A ), so we selected sh-1# and sh-2# for follow-up experiments, and established Hsp27 stable and low expression cell lines Tca8113 / PYM-sh-1# and Tca8113 / PYM-sh-2# and the control cell line Tca8113 / PYM- sh-con, using MTS assay to detect drug sensitivity, found that after interfering with the expression of Hsp27, the sensitivity of Tca8113 / PYM cells to PYM and cDDP increased ( Figure 2A ); at the same time, in order to observe whether the secreted Hsp27 is involved in the chemotherapy resistance of tongue cancer cells, treatment of Tca8113 / PYM cells with Hsp27 neutralizing antibody can also increase the chemotherapy sensitivity (...

Embodiment 3

[0024] Example 3: In vivo experiment of Hsp27 increasing chemotherapy resistance of tongue cancer cells

[0025] The above experiments at the cell level in vitro showed that Hsp27 can participate in the chemotherapy resistance of tongue cancer cells. Here, we further verified the role of Hsp27 in chemotherapy of tongue cancer in nude mice. Tca8113 / pLEX-con and Tca8113 / pLEX-Hsp27 cells After inoculation in the anterior armpit of nude mice to form tumors, from the 15th day after inoculation, the same volume of normal saline (0.2ml), PYM (30mg / kg) and cDDP (3mg / kg) were injected intraperitoneally, once every 3 days , treated 10 times in total, measured the long diameter and wide diameter of the tumor every 3 days to calculate the tumor volume, and drew the tumor growth curve. After the experiment, the nude mice were killed by neck dislocation, the tumor was stripped, and the final volume was measured and weighed. The results showed that the overexpression of Hsp27 could increase ...

technical field

[0001] The invention belongs to the field of biotechnology or medicine, and more specifically, the invention relates to the application of Hsp27 in the diagnosis and treatment of tongue cancer chemotherapy resistance. Background technique

[0002] Heat shock proteins are a group of highly conserved proteins in evolution. In addition to high fever, ischemia, tissue damage, virus or bacterial infection, etc. are also produced, and they have been proved to play various important functions in many fields. Previous studies on heat shock protein 27 (Heat shock protein 27, Hsp27) mainly focused on the anti-apoptosis and anti-oxidative free radical effects of Hsp27 (Small heat shock proteins and protection against injury. Ann N Y AcadSci. 199930; 874 : 66-8.Review); At present, there is no understanding and in-depth research on the application of Hsp27 in the diagnosis and treatment of tongue cancer chemotherapy resistance. Contents of the invention

[0003] In vi...

Examples

Embodiment 1

[0020] Example 1: Chemotherapy can induce the increase of Hsp27 in tongue cancer cells and serum of patients

[0021] The results showed that the level of Hsp27 protein in drug-resistant cells Tca8113 / PYM was significantly higher than that in normal oral mucosa cells Hacat and Tca8113, SCC-25, SCC-15, SCC-9 and CAL-27, which were relatively sensitive to chemotherapy ( Figure 1A ), and the Hsp27 protein level in the drug-resistant cell culture medium was also significantly higher than that of Hacat, Tca8113, SCC-25, SCC-15, SCC-9 and CAL-27 cell culture medium ( Figure 1A ); we treated the above cells with PYM (80mg / L) and cDDP (5mg / L) respectively, and found that chemotherapy could induce Hsp27 protein levels in each cell and in the cell culture medium to varying degrees ( Figure 1A ); In addition, we detected the protein level of Hsp27 in the serum of 30 cases of normal healthy people and 50 cases of tongue cancer patients. Serum of patients with tongue cancer found that ...

Embodiment 2

[0022] Example 2: In vitro experiment of Hsp27 increasing chemotherapy resistance of tongue cancer cells

[0023] In order to observe the role of Hsp27 in chemotherapy resistance of tongue cancer, we silenced the expression of Hsp27 in Tca8113 / PYM cells by shRNA technology, and the interference effects of sh-1# and sh-2# were the most obvious ( Figure 2A ), so we selected sh-1# and sh-2# for follow-up experiments, and established Hsp27 stable and low expression cell lines Tca8113 / PYM-sh-1# and Tca8113 / PYM-sh-2# and the control cell line Tca8113 / PYM- sh-con, using MTS assay to detect drug sensitivity, found that after interfering with the expression of Hsp27, the sensitivity of Tca8113 / PYM cells to PYM and cDDP increased ( Figure 2A ); at the same time, in order to observe whether the secreted Hsp27 is involved in the chemotherapy resistance of tongue cancer cells, treatment of Tca8113 / PYM cells with Hsp27 neutralizing antibody can also increase the chemotherapy sensitivity (...

Embodiment 3

[0024] Example 3: In vivo experiment of Hsp27 increasing chemotherapy resistance of tongue cancer cells

[0025] The above experiments at the cell level in vitro showed that Hsp27 can participate in the chemotherapy resistance of tongue cancer cells. Here, we further verified the role of Hsp27 in chemotherapy of tongue cancer in nude mice. Tca8113 / pLEX-con and Tca8113 / pLEX-Hsp27 cells After inoculation in the anterior armpit of nude mice to form tumors, from the 15th day after inoculation, the same volume of normal saline (0.2ml), PYM (30mg / kg) and cDDP (3mg / kg) were injected intraperitoneally, once every 3 days , treated 10 times in total, measured the long diameter and wide diameter of the tumor every 3 days to calculate the tumor volume, and drew the tumor growth curve. After the experiment, the nude mice were killed by neck dislocation, the tumor was stripped, and the final volume was measured and weighed. The results showed that the overexpression of Hsp27 could increase ...

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